Immunohistochemical study of biopsy material. Igh research. Preparing for the study


[12-027 ] Immunohistochemical study of clinical material (using 1 antibody)

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The study of tissue cells, which is carried out using special reagents.

Synonyms Russian

IHC, immunohistochemistry, immunohistochemical analysis.

English synonyms

Immunohistochemistry, IHC, immunohistochemical analysis.

Research method

Immunohistochemical method.

What biomaterial can be used for research?

Tissue sample/tissue sample in paraffin block.

How to properly prepare for research?

  • No special preparation is required.

General information about the study

Immunohistochemical examination of clinical material is carried out in order to identify the desired substances in it. Finished histological preparations are processed with special reagents containing labeled specific antibodies to the detected substance, which in this situation serves as an antigen. If the desired substance is in the material being studied, then the antibody binds to certain areas of it. As a result, a complex is formed between them and the tissue is stained.

This method is used in the diagnosis of various pathological conditions, its role is especially great in the field of oncology. Immunohistochemical (IHC) research helps not only to diagnose a tumor, determine its nosological variant, identify the primary tumor focus and detect cancerous degeneration of cells, but also to predict the course of the disease and the success of treatment. Factors influencing the prognosis include estrogen and progesterone receptors, Ki-67 (tumor activity marker), her-2 neu (epidermal growth factor, determines tumor sensitivity to the chemotherapy drug trastuzumab/Herceptin), VEGF (vascular growth factor), Bcl-2, p53, etc. Unlike immunohistochemical analysis, they cannot be determined by conventional histological examination.

This method has also found application in the diagnosis of systemic connective tissue diseases and kidney diseases; it allows one to identify bacteria and viruses in the tissues being studied, for example, the Epstein-Barr virus, papilloma virus, Helicobacter pylori(in patients with chronic gastritis), mycobacteria in the diagnosis of tuberculosis, etc.

Based on the IHC analysis, the doctor receives a conclusion with positive and negative markers, and this helps to confirm or refute the hypothesis of other clinical and laboratory studies.

Cervical cancer is one of the most common types of cancer in women. Epidemiological and laboratory data support the role of human papillomavirus (HPV) as the triggering agent for the vast majority of precancerous and malignant lesions of the cervical mucosa. However, HPV DNA can be detected in 95-100% of all cases.

Human papillomaviruses are small, circular, double-stranded deoxyribonucleic acid (DNA) viruses belonging to the family Papillomaviridae. More than 130 are known HPV types. They are classified by risk based on their association with cervical carcinoma. HPV 16 and 18 are the most common types high risk. Invading epithelial cells, the virus causes gene defects in them, thereby promoting tumor development. The expression of p16 and p18 proteins in HPV-infected cells can be easily detected using HPV, and their positivity is highly correlated with HPV positivity. This helps to distinguish between non-tumor dysplasia (as a result of inflammation) and tumor dysplasia (caused by the papilloma virus and can develop into cancer), and also allows for differential diagnosis of moderate and moderate dysplasia. high degree severity (CIN II/III) and initial forms cervical cancer, identifying lesions prone to progression and development of invasive carcinoma. In moderate and high-grade dysplasias (in 80-100% of cases of CIN II and almost all cases of CIN III) and invasive cervical cancer, increased expression of the p16 protein is detected, and it increases as the severity of the lesion increases.

In women with low-grade squamous intraepithelial lesions, focal and diffuse expression indicates disease progression. The absence or focal expression of p16 in high-grade lesions may serve as an additional sign of a favorable course of the pathological process.

Immunohistochemical examination is also used in differential diagnosis.

Celiac disease is systemic disease, caused by genetic intolerance to gluten or the corresponding prolamines found in cereal seeds. Characterized by this pathology damage to the mucous membrane small intestine(development of atrophic enteropathy), the appearance of specific antibodies in the blood serum and a wide range of gluten-dependent clinical manifestations. The disease occurs twice as often among women as among men, and can first appear both in childhood and in adults. Celiac disease can occur with a pronounced or blurred clinical picture, as well as without gastroenterological symptoms. Most often, an erased version of the disease occurs, so diagnosing celiac enteropathy (GE) is difficult.
At primary laboratory stage A serological study is carried out to determine specific biomarkers in the blood. Persons with positive results are shown endoscopy of the small intestine with a biopsy and subsequent histological and IHC analysis of the material taken.

It is known that celiac disease is accompanied by an increase in the number of lymphocytes within the epithelial cells of the small intestine. A distinctive feature is that most intraepithial lymphocytes (IELs) carry a specific T-cell receptor (CD3 γ and CD3δ-positive cells) on their surface. The more active the disease, the more IEL. This feature is used in immunohistochemical studies to determine the predominant type of lymphocytes. This analysis is especially important when there is a discrepancy between the data serological study and histological evaluation on standard areas.

What is the research used for?

  • To determine the type and subtype of the tumor, the degree of its malignancy and the prevalence of the oncological process;
  • for differential diagnosis of proliferative diseases;
  • for determining etiological factor which caused changes in cells;
  • to select effective therapy;
  • to assess the proliferative activity of tumor cells;
  • to assess the effectiveness of treatment;
  • for diagnosing receptor status in cancer (for example, breast cancer, prostate cancer);
  • to determine the primary tumor site;
  • to identify the malignant potential of affected cells;
  • for the initial selection of patients for dispensary registration and further examination;

When is the study scheduled?

  • In the differential diagnosis of dysplasia (high and moderate degree) and the initial stage of cervical cancer;
  • for breast, stomach, prostate cancer in order to determine sensitivity to various types of therapy;
  • when determining the degree of malignancy and assessing the prognosis of the disease;
  • when assessing the effectiveness of therapy;
  • when searching for the source of metastases;
  • in the diagnosis of gastrointestinal stromal tumors (GIST) using an expanded antibody panel including DOG1;
  • in the diagnosis of neuroendocrine tumors;
  • in the differential diagnosis of celiac disease, accompanied by an unclear histological picture;
  • if there is a discrepancy between the results of serological testing and histological evaluation.

What do the results mean?

Reference values

The range of values ​​is individual for each disease and depends on the antibodies used.

The transcript of the study results contains information for the attending physician and is not a diagnosis.

A morphological assessment of the drug is carried out (tumors are classified according to WHO), a descriptive answer is given indicating the assessment of antibody expression. ICG should always be used as a complement to morphological examination, the results should not be interpreted in isolation.

What can influence the result?

For some diseases, there are factors that can influence the outcome. Thus, when diagnosing celiac disease, prescribing a gluten-free diet before the test may lead to a false negative result.



Currently, high-quality diagnostics of many oncological diseases impossible without an immunohistochemical study. This is a highly accurate and informative method that allows not only to identify a tumor, but also to develop optimal treatment tactics.

IHC is mandatory for cancer lymphatic system when lymph nodes or internal organs are affected

IHC studies - what is it?

IHC is a technique used in pathological anatomy to diagnose cancerous and benign tumors. It is prescribed when it is impossible to establish an accurate diagnosis using a standard histological method or when it is necessary to detail the clinically significant parameters of tumors at the molecular level.

Very often histology is not enough. In most cases, the reason for this is the extremely atypical structure of the pathological formation, which significantly complicates making an accurate diagnosis. In particular, for lymphoma and leukemia, IHC is almost always done.

It should be noted that in total there are about 70 types of leukemia and lymphoma. Some of them can be verified only through several studies - histological, immunohistochemical and molecular genetic.

Another diagnosis – “small round cell tumor” – can mean 13 malignant tumors with different courses and prognosis. In addition, each of them requires the development of a special chemotherapy and radiotherapy regimen. The only method that helps differentiate such tumors is immunohistochemistry.

The essence of the technique

With the development of any tumor process proteins foreign to the body are formed - antigens, in contrast to which the human immune system produces antibodies - immunoglobulins. They attach to antigens and bind to epitopes, the main parts of antigenic macromolecules. Antibodies perform two functions at once: binding and effector. Simply put, they directly prevent antigens from causing harm and at the same time activate complement, triggering an immune response.

The role of antigens in in this case belongs to atypical tumor cells. Before immunohistochemical research is carried out, sections of biometal are marked with specific antibodies to them. For further visualization, these antibodies are stained with enzymes. Next, using high-precision optics, the behavior of the test cells is observed.

If protein compounds labeled as antibodies bind to tumor cells, a glow will be visible - fluorescence, indicating the presence of the desired substances. This is how, for example, hormone receptors and tumor markers are detected. If breast cancer is suspected, receptors for estrogen and progesterone are detected in this way.

To whom is it shown?

The main indications for the use of immunohistochemistry are malignant neoplasms. In oncology, this method is used to search for metastases and pathological microorganisms, determine the type and location of the tumor, and also to assess the activity of the pathological process. Using IHC, it is possible to establish a final or, less commonly, intermediate diagnosis for skin cancer (melanoma), sarcoma, lymphogranulomatosis, lymphocytic leukemia and leukemia, and to type the degree of malignancy of the process. The latter is extremely important for neuroendocrine formations, which are also called “hidden killers” due to the fact that they are very difficult to recognize in the early stages.


Immunohistochemical testing is often performed to avoid prescribing aggressive treatment toxic drugs

In some cases, immunohistochemical analysis makes it possible to determine the source of metastases when the localization of the primary lesion is unknown, as well as to carry out differential diagnosis for several neoplasms of different origins.

IHC can be prescribed for infertility, chronic pathologies endometrium, uterus and ovaries, habitual miscarriages. It is also advisable to do it if pregnancy does not occur after several IVF procedures.

Immunohistochemistry will help detect the presence of cells that interfere with conception and determine further tactics for treating infertility.

Immunohistochemical (IHC) study is a method for identifying specific antigenic properties malignant tumors. are used to detect the localization of a particular cellular or tissue component (antigen) in situ by binding it to labeled antibodies and are an integral part of modern cancer diagnostics, providing detection of the localization in tissues of various cells, hormones and their receptors, enzymes, immunoglobulins, cell components and individual genes.

Goals of the IHC study

IHC studies allow:

1) carry out histogenetic diagnosis of tumors;

2) determine the nosological variant of the tumor;

3) identify the primary tumor by metastasis with an unknown primary focus;

4) determine the prognosis of a tumor disease;

5) determine malignant transformation of cells;

6) identify opportunities;

7) identify both resistance and sensitivity of tumor cells to chemotherapeutic drugs;

8) determine the sensitivity of tumor cells to radiation therapy.

How is an IHC study performed?

IHC research begins with the collection of material. To do this, a procedure is carried out, in which a column of tissue is taken from the tumor and nearby tissues, or the material comes from surgery. Then the material is fixed. After fixing, the material is sent to the wiring, which allows it to be prepared for work (degreased and additionally fixed). After processing, all samples are embedded in paraffin to obtain histological blocks. Paraffin blocks are stored forever, so you can conduct an IHC study if you have previously made paraffin blocks.

The next stage of IHC research is microtomy - the laboratory assistant makes sections from paraffin blocks up to 1.0 microns thick and places them on special histological glasses.

Then, routine staining and immunohistochemical examination are carried out sequentially, allowing the phenotype and nosology of the tumor to be more and more differentiated at each stage.

As you can see, IHC research is a complex multi-stage process, and therefore, to conduct IHC research, you should choose the most modern laboratory with highly qualified specialists and a high degree of automation - this way you will eliminate the risks of receiving poor-quality diagnostics. Such a laboratory today is UNIM.

Separately, it should be said about the timing of this study. On average in Russia, IHC studies are carried out within a period of 10 days to several weeks. When you contact UNIM, you can do an IHC study in just 3 days! Also, the advantage of conducting IHC research at UNIM is that you have your research materials from any city in Russia. If necessary, check the cost by submitting an application for research, or call the hotline (toll-free in Russia): 8 800 555 92 67.

Let's first look at what an immunohistochemical study is. By it is meant microscopic examination tissue, which is based on the detection of antibodies to pathological substances.

There are two research methods:

  • direct (direct reaction of antibodies directly to a pathological substance);
  • indirect method(pathological substances recognized by secondary antibodies).

Marking methods:

  • enzymes are protein molecules, also known as RNA molecules. For example: alkaline phosphatase (hydrolase enzyme);
  • fluorescent - the same physical process, for example - fluorescein - an organic compound;
  • electron-dense particles, such as gold.

Use for:

  • study and research of secretory processes, including synthetic ones;
  • hormone receptor recognition;
  • identifying different types and genera of cells based on their properties.

Why is immunohistochemistry used for breast cancer?

In the field of breast examination, this study is used as a preliminary diagnosis to establish an accurate diagnosis, including for malignant tumors. This study makes it possible to determine the following:

  • Accurate formation (benign or malignant) and prescribing the correct treatment.
  • Exact location (presence of metastases in the body and their location).
  • The initial focus of the disease.
  • Stage of disease (for example, grade of cancer).
  • Cell proliferation (recognize the rate of reproduction and development cancer cells).
  • The response of cancer cells to medications, their sensitivity, and sensitivity to chemotherapy and radiation therapy can also be monitored.

The study is able to study the disease in detail, which makes it possible to fully diagnose the disease and select the correct treatment. This plays an important role in the diagnosis, treatment, and life expectancy of cancer patients. We can also say that using this method you can:

Indications for the study

Basically, this diagnostic is used to study (research) any tissue of the human body. The basis for this study is any neoplasm that suggests the development of cancer. The study is also carried out to study the endometrium:

  • with metastases;
  • for infertility;
  • after unsuccessful IVF attempts;
  • for various diseases of the uterus;
  • for various types of pathologies of the pelvic organs;
  • with persistent miscarriage;
  • for endometrial diseases.

The study has no absolute contraindications. The only obstacle to IHC can be the impossibility for some reason of collecting material for research.

How the research works

This type of research can only be carried out in specialized laboratories, and the doctor must have special qualifications and undergo special training.

The sample is taken directly from the affected area of ​​skin using a biopsy. Also, material collection can occur during breast surgery.

Then the resulting material is specially processed and preserved. To do this, it is filled with paraffin. In this form, in paraffin, the material can be properly stored for a long time.

The next step is microtomy. The material is cut into thin layers using a special apparatus.

Subsequently, the cut layers are stained with certain antibodies and all this is studied under a microscope. In some laboratories, colored material is studied using an automatic device.

ICHG Endometrium

Indications:

  • two or three unsuccessful IVF procedures;
  • not the first (permanent) miscarriage early stages;
  • infertility.

During the study, diseases such as:

  • endometritis,
  • hyperplasia,
  • incomplete transformation of the endometrium,
  • developmental phase disorder
  • and other diseases.

As practice shows, diseases associated with the endometrium are 73% the cause of infertility.

Preparing for the study

This study is carried out on a specific day menstrual cycle, usually on days 5-7 they look at the inflammatory process of the endometrium, on days 20-24 of the cycle - secretion is assessed, receptor function is looked at.

  • do not take hormones a week before the test;
  • do not take hemostatic drugs;
  • proper intimate hygiene.

How does the procedure work?

The procedure takes place on a gynecological chair. The woman is anesthetized with special drugs and a speculum is inserted. The genital organs are sanitized. A special device is introduced - a hysteroscope system - and a sampling is carried out. Next, the device is removed, performing organ sanitation.

Then the woman is under the supervision of a doctor until complete recovery, approximately 1-2 hours.

Results take approximately 3-5 days to prepare. Only a specialist can evaluate the results.

Breast ICG results

What does the analysis determine:

HER2neu or, in other words, human epidermal growth factor receptor. Responsible for protein production. When cancer cells are damaged, disorderly growth increases.

If the result is written 1 (+), then this indicates that there is no excess protein, which means the tumor is in poor condition.

At 2 (++), a repeat study is usually prescribed.

If 3 (+++) is considered a positive tumor, which is usually subjected to drug treatment.

ER uPR this type of hormone helps stimulate the growth of pathological tissues. If these indicators are increased, this indicates proper treatment that the tumor responds to hormone therapy. Therefore, if these indicators are reduced, then we can talk about incorrect selection of medications. The tumor develops further.

Ki - 67, this indicator is used to evaluate the stage of the tumor and its characteristics. If the result gives false numbers, then this indicates a rapid progression of the disease, the presence of metastases and, unfortunately, not a good prognosis at all. This result is expressed as a percentage. For example:

  • 11% - quite good prognosis (recovery 93%);
  • 21% - they talk about it as 50% to 50%, it all depends on the human body;
  • all indicators above 30% are assessed as a severe stage - highly aggressive;
  • 90% are incurable and lead to death.

This index also helps to determine the choice of treatment.

The p53 gene helps prevent the growth of pathological tissue. This gene is a barrier to the disease. The onset of the disease is a gene mutation, for example, heredity or a disorder of the protein matrix.

VEGF - refers to a protein that has the function of controlling the vital activity of tissue under conditions of insufficient blood circulation. High levels of this protein indicate the growth of tumor tissue, as a result of “good” nutrition of the tissue.

There are mainly 4 types of cancer:

  • Luminal A - in this case: to female hormones receptors are positive, HER2-negative, ki- 65-13% less.
  • Luminal B - in this case: receptors for female hormones are positive, for HER2 - negative, the ki indicator is 65-15% less.
  • Erb - B2 - in this case: the reaction of receptors to progesterone (estrogen) is negative, to HER2 - positive.
  • Basal-like - negative in all categories.

Determination of PD-1, PDL-1 and PDL-2 protein expression

It is generally accepted that PD-1, PDL-1 and PDL-2 are responsible for the tumor itself. But they may not be present in all pathological tissues. To accurately determine and prescribe the correct therapy, all patients who are directly indicated for immunotherapy undergo a study such as the expression of PD-1, PDL-1 and PDL-2 proteins.

This study is carried out using fluorescence FISH hybridization. As a result of the presence of expression, immunotherapy is prescribed with drugs such as: Nivolumab, Atezolizumab, Pembrolizumab.

Price of immunohistochemical study

Immunohistochemical research is a very complex analysis that uses multiple tumor markers.

The cost of the study depends on the number of factors being tested. The price of the study starts from 4 thousand rubles and up to 20 thousand rubles.

Video: Immunohistochemical studies

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Immunohistochemical study: interpretation and features of the procedure

The most important point in diagnosing cancer is immunohistochemical testing. Every day, microorganisms enter the human body that can trigger the development of a pathological process. The defenses counter this by forming antibodies. This reaction formed the basis for the creation of the IHC study.

The essence of the method

This method of diagnosing cancer is the most modern and reliable. During the development of the tumor process, proteins foreign to the body are formed - antigens. At the same time, the immune system produces antibodies, the main objective which is to prevent the proliferation of pathogenic microorganisms.

The goal of immunohistochemical research is the timely detection of cancer cells. To do this, the patient's biological material is treated with a variety of antibodies, after which it is carefully examined under a microscope. If these protein compounds bind to tumor cells, their glow will be visualized. The appearance of the fluorescence effect indicates the presence of cancer cells in the body.

Today, specialists performing IHC studies have almost all antibodies to various types of tumors at their disposal, which is the key to obtaining reliable results.

Possibilities

The modern type of diagnostics allows you to determine:

  • spread of the tumor process;
  • growth rate of malignant neoplasms;
  • type of tumor;
  • source of metastases;
  • level of malignancy.

In addition, immunohistochemical studies can evaluate the effectiveness of cancer treatment.

Indications and contraindications

By using this method It is possible to study any tissue of the human body. The main reason for prescribing an immunohistochemical study is the suspicion of the presence of a malignant tumor.

In this case the method is used for:

  • determining the type of tumor and the area of ​​its localization;
  • detection of metastases;
  • assessing the activity of the tumor process;
  • detection of pathological microorganisms.

The analysis is also effective for problems with conception.

Immunohistochemical examination of the endometrium is indicated for:

  • infertility;
  • diseases of the uterus;
  • the presence of pathologies in organs reproductive system;
  • miscarriage;
  • chronic endometrial diseases.

In addition, an IHC study is prescribed for patients who do not become pregnant even after several attempts at in vitro fertilization. The method allows you to determine whether there are cells in the body that reduce the likelihood of conception.

There are no contraindications to IHC testing. The only factor that makes it impossible to conduct an analysis is the insurmountable difficulty in collecting the patient’s biomaterial.

How is it carried out?

First, a biopsy is used to obtain a tissue sample from the patient. Less often, material is collected during the process endoscopic examination or surgery. The method of obtaining the sample depends on the type of tumor and its location.

An important nuance is that the collection of material during initial examination must be carried out before starting treatment. Otherwise, the results of the study may be distorted.

After collecting the biomaterial, it is placed in formaldehyde and sent to the laboratory, where it undergoes the following processing:

  1. The tissue sample is degreased and embedded in paraffin. In this form, biological material can be stored for a very long time, due to which the IHC study can be repeated.
  2. Several thin sections are collected from the sample and placed on special slides.
  3. On them, the biomaterial is stained with solutions of various antibodies. At this stage, either a small panel or a large one can be used. In the first case, reactions are studied after using 5 types of antibodies, in the second - up to several dozen.
  4. During an immunohistochemical study of cancer of any organ, a fluorescence effect appears, which allows a specialist to determine the type of malignant cells.


Interpretation of results

As a rule, the conclusion is ready in 7–15 days. The period depends on the type of panel used (small or large). The advanced method takes longer.

The study of sections of biomaterial is carried out by a pathologist who has the knowledge and skills (confirmed by an official document) necessary to carry out the analysis.

When interpreting the results, special attention is paid to the Ki-67 indicator. It is he who provides information about the degree of malignancy of the process. For example, if the result of an immunohistochemical study for breast cancer is no more than 15%, the prognosis is considered to be more than favorable. A level of 30% indicates the activity of the tumor process, i.e. about the rapid speed of its development. As a rule, it stops after a course of chemotherapy.

According to some statistics, if Ki-67 is less than 10%, the outcome of the disease will be favorable (in 95% of cases). A mark of 90% or higher means almost 100% fatality.

In addition to the malignancy indicator, the conclusion indicates:

  • antibodies to which similarities have been identified (tropism);
  • type of cancer cells, their quantitative significance.

It is important to understand that an accurate diagnosis is made after receiving and studying the information collected through all the diagnostic procedures. Although IHC analysis is considered the most informative method compared to histology, sometimes it is necessary to use both methods. The immunohistochemical study is deciphered exclusively by the oncologist.

Finally

In modern medicine, special attention is paid to the diagnosis of cancer. The most modern and informative method is immunohistochemical research. With its help, not only the presence of cancer cells is detected, but also their type and the rate of development of the malignant process are determined. In addition, based on the results, the effectiveness of the prescribed treatment is assessed.

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IHC examination of the breast - transcript

Immunohistochemical study (IHC) is a method for studying glandular tissue of the breast, which uses a special reagent to obtain full description cells:

  • establish the origin of tumor cells;
  • determine its structure;
  • diagnose primary formation based on existing metastases;
  • accurately calculate the duration of the disease, the age of the tumor;
  • determine the correct treatment method.

IHC analysis of the mammary gland is prescribed both when an oncological process is suspected and during its course, in order to diagnose the effectiveness of chemotherapy treatment.

What can IHC determine?

To begin with, it must be said that deciphering the result of an IHC study of the mammary gland should be done exclusively by a doctor. Only he, knowing fully the features of the course of the disease, can interpret the results obtained.

IHC performed in breast cancer determines the nature of the tumor. The most common receptors used in breast IHC are:

  • estrogen (ER);
  • progesterone (PR).

It has been established that a tumor containing a large number of of these receptors behaves non-aggressively and is inactive. When treating this form, hormonal therapy is significantly effective. Favorable prognosis in 75% of cases.

When interpreting the results of breast IHC analysis, percentage units of measurement are used. In this case, the ratio of the number of cells with expression (susceptibility) to estrogens and progesterone is determined, total number tumor cells. In this case, the result is displayed as the ratio of the number of nuclei of stained cells to unstained ones, in total to 100 cells.

Due to the complexity of such calculations and their interpretation, the assessment of the result is carried out exclusively by specialists.

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Immunohistochemical study for breast cancer

If a patient is suspected of having a malignant tumor inside the mammary gland, the doctor must prescribe comprehensive study diseased area of ​​the organ. Some of the types of examination are standard and form the basis of the medical protocol when detecting oncology in the chest. Not every tumor found in the breast can be immediately diagnosed as malignant. To make such a conclusion, the oncologist is obliged to send the woman for an immunohistochemical study for breast cancer.

Only after cancer cells that have pathogenic properties and are capable of actively dividing and spreading beyond the mammary gland are detected in the biological material of the tumor does the neoplasm receive the status of malignant.

How does immunohistochemical examination of the breast occur?

After a new formation is detected in the mammary gland, one of the mandatory tests to perform is immunohistochemistry. The essence of its implementation is that doctors perform a minor surgical operation.

A micro-incision is made on the diseased breast, into which special equipment is inserted to remove part of the tumor tissue. The patient is under anesthesia. After obtaining the biological material of the neoplasm, it is treated with specific antibodies. A malignant tumor produces its own aggressive agents, which have a protein structure.

They are also endowed with the ability to interact with others biological substances, bind them, and form a single fabric with them. Based on the results of processing the tumor material with antigens, the tissue is placed under a microscope for detailed visual examination.

Fragment benign tumor after treatment with antibodies, it does not change its color, and the cellular structure of the biopath and antibodies remains unchanged, there is no active contact between them. Malignant cells behave completely differently, and this is clearly visible through the microscope lens.

Breast cancer is the most common cancer in women and the second leading cause of cancer-related death. Early diagnosis, timely and correct treatment can significantly increase the chances of recovery. Traditional immunohistochemistry (IHC) techniques work with very small tissue samples. This circumstance, combined with the use of antibodies specific to tumor cell antigens, makes this method an effective tool in the hands of a pathologist involved in the diagnosis and prognosis of the course of cancer.

  • Estrogen Receptor
  • Progesterone Receptor
  • HER-2/neu
  • Ki-67
  • p120 Catenin
  • CadherinE
  • The “gold standard” for diagnosing breast tumors is the hormonal profile PR, ER, HER-2/neu, Ki-67- this is the diagnosis of all available receptors that are responsible for the activity of cancer tissue. Includes the study of several indicators.
  • PR, ER- these are specific receptor proteins that respond to the production of estrogen and progesterone. Most breast cancers (about 80 percent) actively respond to changes in hormone levels. Determining the reactivity of these receptors plays a critical role in assessing the potential for hormone therapy.
  • HER-2/neu is a gene protein structure that is located in cancer tissue. It is a receptor that responds to the production of specific antibodies. It is advisable to study this parameter from the point of view of determining the prognosis of cancer treatment. With high HER-2/neu activity, the tumor is difficult to treat; monoclonal therapy is first required, aimed at suppressing the activity of this structure.
  • Ki-67 is a protein structure that has the ability to become activated during active tumor growth. The study of this indicator makes it possible to assess the prognosis for the patient’s life. The higher the expression characteristics of Ki-67, the less tumor differentiation, the less chance a sick woman has of recovery.

Prostate tumors

Prostate cancer is one of the most common cancers in the world. Most cases (50 – 70%) are diagnosed at stages 3-4, including 25% with generalization of the tumor process. Unfortunately, early diagnosis of cancer is difficult due to the frequent absence of characteristic symptoms. Along with clinical methods, the most informative method is the histological examination of prostate biopsies.

Main diagnostic markers:

  • p63;
  • PSAP (ProstaticAcidPhosphatase);
  • PSA (Prostate Specific Antigen);
  • P504s (= AMACR - Alpha methylacyl-CoA-racemase);
  • Cytokeratin High Molecular Weight (34betaE12);
  • ERG (ETS Related Gene);
  • PSMA (Prostate Specific Membrane Antigen);
  • Androgen Receptor;
  • Bcl-X;
  • Cytokeratin 5 & 6;
  • Cytokeratin Pan;
  • Keratin 8;
  • Cytokeratin 8 & 18;
  • Ki-67;
  • p53;
  • Synaptophysin;
  • Basal Cell Cocktail - Cytokeratin HMW + p63.

Lung tumors

Lung cancer is one of the most common causes of death. Around the world, about 1 million people die from this disease every year. In men, lung cancer in 85-90% of cases is associated with smoking. The prognosis for lung cancer remains unfavorable. Without treatment, up to 90% of patients die within 2 years from the moment of diagnosis. With surgical treatment, the 5-year survival rate is about 30%. Surgery combined with radiation and drug therapy increases 5-year survival by 40%. The presence of metastases significantly worsens the prognosis. Modern diagnosis and treatment of patients with lung cancer cannot do without morphological verification of the tumor with clarification of the histological structure and degree of anaplasia (differentiation) of tumor cells. The immunohistochemical method remains one of the most informative methods at this stage of diagnosis

Main diagnostic markers:

  • Thyroid Transcription Factor-1;
  • Cytokeratin 7;
  • NON-SMALL CELL LUNG CANCER
    • ALK (DE5F3);
  • SQUAMOUS CELL LUNG CANCER
    • Cytokeratin 14;
    • Cytokeratin 5/6;
    • EGFR;
  • SMALL CELL LUNG CARCINOMA
    • ChromograninA;
    • Synaptophysin;
  • ADENOCARCINOMA OF THE LUNG
    • Cytokeratin Pan.

Melanoma

Melanoma (lat. melanoma, melanoma malignum from ancient Greek μέλας - “black”) (oral Melanoblastoma) - a malignant tumor developing from melanocytes - pigment cells, producing melanin. One of three varieties skin cancer, and the most dangerous of them all. Mainly localized in the skin, less often - in the retina of the eye, mucous membranes (oral cavity, vagina, rectum). One of the most dangerous human malignant tumors, often recurrent and metastasizing by lymphogenous and hematogenous routes to almost all organs. Verification of skin melanoma and its metastases remains one of the most difficult tasks for an oncomorphologist. Among non-pigmented melanomas there are nodular, superficial spreading, lentigo type, clear cell, spindle cell, pleomorphic, small cell, myxoid, “nevoid”, signet cell and other forms.

Main diagnostic markers:

  • MelanomaAssociated Antigen (MAA);
  • CD63;
  • Melanoma Marker (HMB45);
  • MART-1/Melan-A;
  • Melanoma (gp100);
  • Tyrosinase;
  • Microphthalmia Transcription Factor (MiTF);
  • Nerve Growth Factor Receptor (NGFR);
  • S100;
  • Melanoma Pan (HMB45 + A103 + T311);
  • MART-1 + Tyrosinase;
  • Vimentin.

A standard panel contains about five immunohistochemical markers:

  • p53- degree of melanoma mitosis activity;
  • Ki-67- assessment of the intensity of proliferation determines the prognosis of the disease;
  • bcl-2- a protein that prevents the natural apoptosis of skin melanoma, the likelihood of metastasis is assessed;
  • HMB-45- assessment of melanocyte function;
  • S-100- a typical antigen found only in melanoma, allowing it to be distinguished from other tumors or benign formations.

Lymphoproliferative diseases

Lymphoma is a malignant tumor disease of the lymphatic system. Lymphomas include lymphogranulomatosis (Hodgkin's lymphoma) and all other types of lymphomas - non-Hodgkin's lymphomas (NHL). Based on the type of lymphoid cells from which the tumor arises, B-, T- and (rarely) NK-cell lymphomas are distinguished. Most lymphomas are B-cell. Diagnosis of lymphoproliferative diseases currently occupies a leading position in modern pathology and requires one of the most extensive panels of markers. The total incidence of all types of non-Hodgkin's lymphoma in European countries is 12-15 cases per 100 thousand population per year. The risk of their occurrence increases with age. Epstein-Barr virus infection is associated with increased risk diseases of various types of lymphomas, including Burkitt's lymphoma. In children, non-Hodgkin's lymphomas are relatively rare: no more than 5% of all cases of NHL occur in childhood and adolescence. However, lymphomas still occupy in the structure malignant diseases childhood third place in frequency - after leukemia and tumors of the central nervous system.

Main diagnostic markers:

Stromal tumors of the gastrointestinal tract (GIT)

GI tract occurs predominantly in the stomach (60%) and small intestine (25%), but is also observed in the rectum (5%), esophagus (5%) and a number of other places (5%), including the appendix, gallbladder, mesentery and omentum. The age of the sick patients ranges from adolescence to 90-year-olds, but most patients are older with a peak around 60 years. Most studies show a slight male predilection. In 1998, it was shown that the GI tract expresses the tyrosine kinase receptor KIT (CD117). The cause of these tumors was found to be interstitial cells of Cajal (ICC). Like the GI tract, Cajal cells express KIT and the majority are positive for CD34. Subsequent studies with a large number different laboratories have confirmed that KIT is the single most specific marker of the gastrointestinal tract. Immune-detectable KIT is present on the cell surface and/or cytoplasm of GIT tumor cells in approximately 90% of cases. In the vast majority of tumors, KIT expression is strong and uniform, but in some cases only focal positive reactivity is demonstrated and KIT is absent in a small subgroup (~5%) of tumors that, according to other morphological and immunophenotypic features, correspond to the GI tract. Among KIT-positive BUT, CD34 expression is detected in 60-70% of cases, while 30-40% have positive reaction on smooth muscle actin (SMA), and 5% on S-100 protein. None of these antigens are specific for the gastrointestinal tract. Desmin expression in true KIT-positive GI tracts is extremely rare (1-2% of cases) and usually focal. This form Oncological diseases are difficult to diagnose morphologically. Using modern panels of markers it is possible to clearly and reliably diagnose different forms of the described pathology. Immunohistochemical examination is mandatory.

Main diagnostic markers:

  • CD117 c-kit;
  • CD34;
  • Desmin;
  • Beta-Catenin;
  • S100;
  • GFAP;
  • CD99;
  • ActinSmoothMuscle.

Colorectal cancer

Colorectal cancer is the third most commonly diagnosed cancer in the United States (excluding skin cancer) among men and women. Colorectal cancer incidence rates have been declining over the past two decades (from 66.3 cases per 100,000 people in 1985 to 45.5 cases in 2006). This is associated with the increased use of colorectal screening tests, which make it possible to detect and remove gastrointestinal polyps before they develop into cancer. In contrast to the overall decline, among young adults under 50 years of age, for whom screening is not recommended due to medium degree risk, the incidence of colorectal cancer has increased by about 2% per year since 1994 in men and women. In 2016, the US death rate from colorectal cancer was 49,500. Mortality rates from colorectal cancer have declined in both groups of men and women over the past few decades, with a steeper decline in the most recent time period. This decline reflects declining morbidity rates and improved early diagnosis and treatment. Early stages of colorectal cancer usually have no symptoms, so screening is often necessary to detect the disease at this early stage. Progression of the disease can cause bleeding from the rectum, the appearance of blood in the stool, changes in bowel movements, and cramping pain in the lower abdomen. The use of IHC in colon cancer is considered at several levels: to characterize tumors (endocrine or epithelial type), hereditary predisposition and for prognosis purposes. The predominant use of IHC is to identify possible or suspected metastases in which the colon is a possible primary. Typical localization for colon metastases are the liver and lungs, both organs that can produce cancer morphology identical to metastases from the colon. IHC, (FDA regulatory class I), is used after initial tumor diagnosis by histopathological examination and is not included as an independent test for clinicians.

Main diagnostic markers:

  • Beta-Catenin;
  • BRAF;
  • CDX-2;
  • COX-2;
  • Cytokeratin 7;
  • Cytokeratin 19;
  • Cytokeratin 20;
  • MLH1;
  • MLH2;
  • MLH6;
  • MSLN;
  • MUC1;
  • MUC2.

Metastatic carcinoma

The most common use of immunohistochemistry in the study of liver tumors is to determine the source of metastasis when the primary location of the tumor is unknown. The development and application of immunostaining panels can help solve almost all diagnostic problems. 2-6 Cytokeratins (CK) 7 and CK 20 are the first step in the identification of many tumors and, with additional immune responses relatively specific to tumors of the female and male reproductive tract, often make it possible to identify the primary site of a metastatic tumor.

Main diagnostic markers:

  • Cytokeratins of various molecular weights (CK 18, CK 19, CK 7, and CK 20, etc.).
  • HER2/neu- is a membrane protein that is encoded by the ERBB2 gene. Increased expression is important in the pathogenesis and progression of certain malignant processes. Testing of this receptor is an important biological marker for cancer of the stomach, breast, uterus and its appendages.;
  • Ki-67- nuclear antigen, consisting of two polypeptide chains and being the main part of the nuclear matrix. Its expression makes it possible to isolate proliferating tumor cells that are in the active phase of the cell life cycle. This marker allows us to determine the phenotype and growth rate of the tumor, the risk of its metastasis, the potential response to treatment measures and the outcome of the pathological process.

Stomach tumors.

Immunohistochemical studies (IHC) are generally not required to evaluate benign and malignant gastric epithelial tumors, since histopathology usually provides diagnosis, but IHC is needed when studying metastatic cancer stomach when the origin of the tumor is unclear, or when the macroscopic/radiological appearance of the tumor is confusing (eg, gastric cancer invades directly and extensively into the liver and is histologically indistinguishable from cholangiocarcinoma). In addition, IHC may be useful in identifying certain types of gastric carcinomas, including hepatoid adenocarcinoma, in which hepatic differentiation can be confirmed by positivity for alpha-fetoprotein AFP. Gastric adenocarcinomas will react with many antibodies against keratins, including CK 18, CK 19, CK 7, and CK 20. When CK 7 and CK 20 are used together, many gastric adenocarcinomas will stain with both CK 7 and CK 20. Approximately 25% cases will have the phenotype CK 7+/CK 20-, or CK 7-/CK 20+), and a small number of cases will be negative for both markers. It was initially thought that CDX-2, a specific marker for colon cancer, would be reactive in more than 50% of cases and may be indicative of a lesser degree of invasiveness. Gastric adenocarcinoma, both the intestinal type and the signet ring cell carcinoma type, may have neuroendocrine differentiation and may not be obvious from the histological picture, but appear by chromogranin and/or synaptophysin staining

Determination of EGFR receptor expression in epithelial tumors

Immunohistochemical (IHC) determination of EGFR receptor expression in colorectal cancer And lung tumors, as well as for tumors of the neck and head, are carried out for an adequate selection of chemotherapy treatment regimens.

EGFR ( Epidermal Growth Factor Receptor) is one of the transmembrane receptors, expressed on the surface of epithelial cells and is involved in the regulation of cell growth and differentiation. Cell division occurs much faster in its presence. When the EGFR receptor is activated after binding to growth factors (EGF and TGF-a), mechanisms are triggered that lead to tumor growth and the proliferation of cancer cells increases, and the process of metastasis is also stimulated. EGFR expression is an indicator indicating that tumor growth is stimulated activity of the epidermal growth factor receptor. Since activation of the receptor occurs due to the substances of the tumor itself, it would be more correct to talk about the expression of EGFR by the tumor. Expression of EGFR is found in the following forms of cancer: lung, neck and head, colon and rectum. Immunohistochemical determination of EGFR expression allows us to determine the status of these receptors and prescribe treatment. The expression of EGFR is directly related to the degree of malignancy and the stage of tumor development. A specialist, in accordance with the data obtained from an immunohistochemical study, classifies the tumor as EGFR-negative or EGFR-positive.

Overexpression of EGFR indicates high malignancy, late tumor development and metastatic processes. This factor is unfavorable with regard to the prognosis of the disease and indicates the high proliferative activity of the tumor, aggressiveness, and resistance to therapy.

A low degree of EGFR expression indicates tumor regression and positive dynamics in treatment.

Immunohistochemical study of receptor sensitivity to estrogen and progesterone in the endometrium

An immunohistochemical study of receptor sensitivity to estrogen and progesterone in the endometrium is carried out to identify the causes of lack of fertility, as well as to assess the malignancy of processes in the uterine tissue. The study is complex, carried out according to plan, and requires the appropriate qualifications of a pathologist.

Estrogen receptors (ER) and progesterone receptors (PR)- these are sensitive markers that respond to fluctuations in certain hormones that affect the growth of tumors and the development of hyperplastic processes in the endometrium. They are located both in the tissues of the uterine epithelium and in the cells of the mammary gland. Their definition makes it possible to assess the impact hormonal factors on the progression of malignant growth, and in addition, identify the presence of other foci of activity in addition to the uterine one. They are included in the mandatory screening program for sick women with suspected infiltrative cancer activity.

Immunohistochemical examination is carried out when:

  • infertility;
  • endometrial tumors;
  • dysfunction of menstruation;
  • hyperplastic processes in the endometrium.

In case of infertility, the technique will allow you to find out whether the fertilized egg can attach to the wall of the uterus. For cancer, the method is not an early diagnosis method. Assessment of receptor activity allows us to identify the presence of metastasis and evaluate the effectiveness of treatment in the uterine cavity. In case of ovulation disorders, the technique determines the effectiveness of hormonal therapy. With changes in the uterine cavity of a hyperplastic nature, increased activity of receptors for estrogen and progesterone indicates the development of low-grade tumors, potentially life-threatening to the woman.

General principle of the result- the greater the expression of receptor activity, the higher the likelihood of tumor progression. The lower the activity, the less possibility of natural fertilization.

Chronic endometritis

Chronic endometritis is called inflammation of the mucous layer of the uterus, which is provoked by various viruses or pathogenic microorganisms. Morphological and functional changes in the endometrium occur in the pathological focus. Chronic endometritis is a clinical and morphological syndrome in which, as a result of damage to the endometrium by an infectious agent, multiple secondary morphofunctional changes occur that disrupt the cyclic biotransformation of the uterine mucosa, which leads to persistent disruption of menstrual and generative functions. Frequency The incidence of chronic endometritis in the population is 2.6-51%. Moreover, among these women, 60.4% are infertile, and unsuccessful attempts at IVF and embryo transfer were noted in 37%. In 2006, the International Federation of Gynecology and Obstetrics equated the concepts of “non-developing pregnancy” and “chronic endometritis”.

Causes of chronic endometritis:

  • infections of the pelvic organs, vagina and cervical canal;
  • intrauterine device;
  • early intimate contacts;
  • radiotherapy of the pelvic organs;
  • surgical intervention into the uterine cavity;
  • alcohol abuse and smoking.

Practitioners include inflammatory processes, occurring in the pelvic organs, to autoimmune pathology. To determine the nature of the disorders and identify patients with a pathological response of the immune system, which provokes inflammation of the endometrium, an immunohistochemical study is prescribed, which is carried out using a standard panel of monoclonal antibodies: CD16, CD20, CD138, CD56, HLA-DR.

Immunohistochemical study of endometrial receptivity (implantation window)

Endometrial receptivity is a complex of structural and functional characteristics of the endometrium, which determines its ability to implant. Since the early 90s of the last century, the concept of “endometrial receptivity” has begun to acquire its modern meaning as a process of complex integration and multi-level “dialogue” between the endometrium and the embryo during a specific period of the “implantation window”. The duration of the “implantation window” in humans is on average 4 days: from the 6th to the 8-10th day after the peak of LH secretion, or 20-24 days of the menstrual cycle (with a 28-day menstrual cycle). Currently, three levels of receptivity are distinguished: genetic, proteomic and histological. When the “implantation window” opens in the endometrium, the expression of 395 genes (ApoE, PLA2) increases and the expression of 186 genes (ITF, various proteases, extracellular matrix proteins, etc.) decreases. Among the proteomic markers associated with endometrial receptivity, various adhesion molecules, growth factors, cytokines and receptors are distinguished: the IL-1 family, LIF and LIF-R, αVβ3, TNF-α, IFN-γ, etc. Of these, leukemia is the most studied -inhibiting factor (LIF) is a member of the IL-6 family. Its maximum expression in the endometrium is observed on the 20th day of the cycle. The third level of receptivity is histological. The “window of implantation” in the endometrium corresponds to the middle stage of the secretion phase of the menstrual cycle. The endometrium can have receptive properties only if molecular markers of receptivity are detected exactly at the middle stage of the secretion phase of the menstrual cycle. One of the key ultrastructural formations involved in the formation of receptivity is pinopodia. These are microscopic protrusions in the apical part of the surface epithelium of the endometrium, formed in place of microvilli in the “implantation window” and protruding into the uterine cavity. It is assumed that the main receptors for attachment of blastocytes are located on the surface of the pinopodia, where LIF is also concentratedly expressed. Any imbalance in the expression of steroid receptors can lead to disruption of the morphofunctional properties of the endometrium and its receptivity. Therefore, determining the level of ER and PR in the middle stage of the secretion phase makes it possible to complement the morphological study of the endometrium and evaluate its receptivity. Normally, the PR/ER ratio in the stroma ranges from 2 to 4. In the middle stage of the secretion phase, a physiological decrease in the level of ERα in the endometrium is observed. This is a critical event that releases certain genes from suppressive influence and provides the signal for the onset of intrauterine receptivity.

Overexpression of ER α in the middle stage of the secretion phase causes a disruption in the expression of biological markers of implantation and disrupts endometrial receptivity.

The comprehensive research program consists of the following panel of antibodies: ER, PgR, CD56, CD138, LIF, as well as counting the number of pinopodia.

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