In suspected tumor education, the patient prescribes a number of standard surveys.

The diagnosis of malignant lesion can be exposed only on the basis of an assessment of changes in bioptate - immunohistochemical study (IHD) is considered one of the most reliable methods for detecting tumor cells.

What is an immunohistochemical study?

The essence of immunohistochemical research is a study under a microscope of samples of biological material, that is, tissues obtained by biopsy.

Pre-tissue is treated with certain specific antibodies.

And the cancer cells themselves are the objects of a comprehensive study for several decades.

During these studies, scientists managed to establish that tumor cells produce specific proteins or as they call them - antigens.

These proteins have the ability to bind to antibodies. It was on this that the principle of IGI was built - the patient-pointed fabric for research is processed by a number of standard antibodies and a study under a microscope is carried out.

If antibodies come into interaction with cancer cells, they have a fluorescence property, that is, there is a glow, characterized by different wavelengths. That is, if a similar change in the explored bioptate is detected, then you can already have more likely to set malignant education.

Today, antibodies are developed and actively used to most of the most common neoplasms.

Immunohistochemical study allows:

• Determine the type of neoplasm and its subspecies.
• Set as far as the primary cancer organism is common.
• Determine the source of metastasis if the biopsytte is obtained from the secondary foci of cancer.
• Assessing how efficient treatment of patients with onco-scabers.
• Install the stage of malignant neoplasm.
• Find out the proliferation of tumors, that is, to establish the growth rate of the neoplasms.

IGH is considered a more informative method compared to more familiar histological. But in some cases it is necessary precisely histology, therefore it is desirable to use both analysis.

Indications

The IGG method can be used to study almost any tissues in the human body. This type of research is appointed mainly if there is a suspicion of the development of the tumor process.

Immunohistochemical research is used:

• To determine the type of primary, mostly single, neoplasms.
• To identify metastases.
• When it is necessary to determine the forecast of the development and flow of tumors.
• As a method for studying receptors to a row of hormones.
• To determine the type of lymphoproliferative states.
• To detect microorganisms.

There are no contraindications to IGOs. This analysis cannot be carried out only if irresistible difficulties occur in the fence of samples of damaged tissues.

How is the analysis?

IGH is carried out in several stages, the very first is a Dolboaton, that is, fence fabrics for analysis.

The sample is obtained by biopsy, in some cases it is possible to pressing a piece of tissues or their withdrawal when conducting an endoscopic or surgical operation.

How will be received biological materialdepends on the localization of the neoplasm and from its type. The folding material is placed in formalin and only after that they are sent to the laboratory center.

In the laboratory, the sample of the fabric is subjected to several changes:

• The material is degreased and poured with paraffin. Thus, histological blocks are obtained, it is possible to store them forever, therefore IGH can repeat if necessary.
• The next stage microtoming is to obtain the finest sections with a width of up to 1.0 μm from paraffin blocks. Slices are distributed on special glasses.
• The resulting sections are painted with immunohistochemical preparations, that is, solutions of antibodies at a certain concentration. A small panel can be used to study - it includes 5 types of antibodies. The large panel contains from 6 to dozens of markers. What antibodies will enter into interaction with tumor cells depends on the alleged type of neoplasm.

The results of the IGH are becoming known for 7-15 days.

IGH with breast cancer

To determine certain immunohistochemical markers were derived. In suspected malignant education, be sure to be the number of such receptors as estrogen and progesterone.

The excess of these hormones provokes the growth of malignant education and affects the appearance of metastasis. IGHs for expression of progesterons and estrogen allowing more accurate to establish the stage of the disease and to find out whether hormonal treatment is shown in this case.

Tumors S. high concentration Most of the hormones are mostly not distinguished by increased activity and are successfully treated with anti-cormonal drugs.

When conducting IGH, it is necessary to determine such an indicator as Ki-67, it indicates the malignancy of the process. If the KI-67 breast cancer rumbles only to 15%, the outcome of the disease is considered to be favorable.

At a level of 30%, they talk about the rapid speed of development of the tumor, the cessation of its growth occurs under the influence of chemotherapy. If the indicator is less than 30%, the treatment of patients is carried out by hormonal means.

The examination of the patients made it possible to establish that if Ki-67 is less than 10%, the survival rate approaches 95%. At the level of this indicator, in 90%, the fatal outcome is observed in almost one hundred percent.

Immunohistochemical research is appointed not only with malignant damage to the breast, this study is informative at:

• Infertility.
• Pathological changes in reproductive function.

IGH Endometry

Immunohistochemical study of endometrial tissues is assigned:

• With infertility.
• With a frequent of non-obscure pregnancy.
• Patients with several unsuccessful attempts eco.
• Diagnosis chronic form Endometrial.

IGH allows you to establish whether there are cells that prevent conception. This analysis simultaneously determines how the uterine tissue receptors are reacting for hormonal stimulation, if the pathology is detected, this indicates changes in endometrial - endometritis, hyperplasia, desynchronization of the endometrium differentiation process.

It is such violations in two cases of three become the main causes of problems with conception and tooling the fetus.

Fence of endometrial fabrics, depending on the detected pathology, is carried out in different days of the cycle, the biopsy day must appoint a doctor. I can install cancer disease and the infection of HPV of several subspecies.

Decoding results

Exploring the prepared samples by the pathologist, he must have a certificate of special preparation of tests on the IGH method.

In conclusion, the indicators of antibodies, to which the pathness of the material under study is determined.

The morphological structure of the tissue is necessarily indicated, that is, the type of atypical cells and their number.

The identification of certain antigens indicates a type of cancer pathology. But it must be borne in mind that the diagnosis is exhibited only on the totality of these diagnostic procedures. Interpret the results of IGH can oncologist.

Research price

The cost of immunohistochemical research depends on how many antibodies are used when analyzing.

Standard study (from 2 and up to 5 antibodies) in most clinics is within 4-5 thousand rubles. If a dozen antigens are revealed immediately, then the cost can be much higher - from 15 thousand rubles.

Video about how immunohistochemistry helps to diagnose a malignant tumor:

Immunohistochemical (IGH) Study is a method for identifying specific antigenic properties of malignant tumors. Used to identify the localization of a cellular or tissue component (antigen) in situ by binding it with labeled antibodies and are an integral part modern diagnosis Cancer, providing localization detection in tissues of various cells, hormones and their receptors, enzymes, immunoglobulins, cell components and individual genes.

Goals of IGG research

IGH studies allow:

1) carry out histogenetic diagnosis of tumors;

2) to determine the nosological version of the neoplasm;

3) to identify the primary tumor in metastase with an unknown primary hearth;

4) determine the prognosis of the tumor disease;

5) to determine the malignant transformation of cells;

6) determine the possibilities;

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

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

How is IGG research?

IGG research begins with the fence of the material. For this, they are carried out at which the tumor tumor and nearby tissues are taken away, or the material comes from the operation. Then the material is fixed. After fixing, the material is sent to the wiring, which allows you to prepare it for work (degreased and fix it). After the wiring, all samples are poured with paraffin, getting histological blocks. Paraffin blocks are stored forever, so you can conduct IGH research in the presence of paraffin blocks made earlier.

The next stage of the IGG studies is microtoming - a laboratory manner makes cuts from paraffin blocks to a thickness of up to 1.0 microns and places them on special histological glasses.

Then there are consistently routine painting and immunohistochemical study, allowing at each stage more and more differentiate the phenotype and nosology of the tumor.

As you can see, IGH research is a complex multi-stage process, and therefore for conducting IGH studies should choose the most modern laboratory with highly qualified specialists and a high proportion of automation - so you will reduce the risks of obtaining poor quality diagnostics. Such a laboratory today is young.

Separately, it should be said about the timing of this study. On average in Russia, IGH research is carried out in time from 10 days to several weeks. When contacting you, you can make the research of the IGG in just 3 days! Also the advantage of the implementation of IGH studies in the Yunima is your materials on a study from any city of Russia. If necessary, check, submitting an application for research, or call hotline (for free in Russia): 8 800 555 92 67.

Definition method Bioptate histological examination according to the histological classification of the World Health Organization (WHO) with staining hematoxylin-eosin. IHG study using the spectrum of tissue-specific and prognostic antibodies (up to five antibodies) (peroxidase and avidin-biotin methods).

The material under study See the description

Available Departure to the house

A comprehensive study of biopsy metastatic formations of various localizations, including a morphological description and evaluation of the expression of the spectrum of tissue markers, to determine the histogenesis of the primary tumor focus.

According to statistics, in 3-15% of cases malignant tumors Manifest with metastases. Metastases without a detected primary focus are characterized by random, atypical localization and rapid progression of the process. The average life expectancy of patients with three and more metastatic lesions of the lesion is three months. At the same time, the localization of the primary focus in 60-70% of patients is detected only on autopsy. Timely IGH diagnosis determines the tactics of treatment before the detection of the primary hearth.

Patomorphological diagnosis of metastases of unbearable tumors provides for the definition of their morphological type, clarifying the likely source of metastasis, an assessment of malignant potential. The first task is solved with a routine histological examination of the biopsy material (staining with hematoxylin-eosin), the second - applying special research methods: histochemical, immunohistochemical (IGH) or molecular-genetic method FISH (FlueScence in Situ Hybridization).

With routine morphological diagnostics, metastatic tumors, according to recommendations European society Medical Oncology ESMO (European Society for Medical Oncology) (2004) is divided into five major categories: adenocarcinoma, flat-cell cancer, neuroendocrine cancer, undifferentiated cancer, undifferentiated tumor. These morphological categories, along with the data of the prevalence of the process, in many cases allow to determine an adequate survey plan (and treatment). So, for example, when identifying a metastase of flat-belling cancer in the cervical lymph nodes Endoscopic examination of the upper respiratory and digestive tract organs is necessary. The IHG study, depending on the morphological type of the neoplasm, allows you to clarify the histogenesis of the tumor and / or determine the likely localization of the primary hearth.

For IHG research primary tumors and their metastases are used wide spectrum Tissue-specific markers: -citospecific (leukocyte differentiation clusters (CD - Clusters of Differentiation), smooth muscle actin, moglobin, thyroglobulin); -Markers proliferation (Ki67, PCNA - PROLIFERTITIN CELL NUCLEAR ANTIGEN); - Tumor markers - oncofetal antigens (fetoprotein, carcinoembrium antigen); -Gurnones (estrogen, progesterone); -Fermines, protein products of cell oncogenes, etc.

Unified algorithms for the IGH studies of metastases of tumors without a refined primary hearth is not developed. The correct determination of the direction of differentiation of tumor cells and a number of biological tumor parameters in itself is an indication for assigning certain patterns of therapy. For example, the detection of adenocarcinoma metastases in the lumphosals of the axillary region may indicate the treatment of therapy similar to the treatment of breast cancer of the appropriate stage. The detection of expression of estrogen receptors and progesterone in such an adenocarcinoma may be indicated to the appointment of anti-humon therapy, regardless of the presence of a determined tumor node in the breast. An example of an algorithm recommended for use in differential diagnostics.

Material for research: tumor bioptat, fixed in paraffin block.

Literature

1. Blaszyk H., Hartmann A., Bjornsson J. Cancer of Unknown Primary: Clinicopathological Correlations. Apmis. 2003; 111 (12): 1089-1094.
2. Dabbs D.J. Diagnostic ImmunoHistochemistry: Theranostic and Genomic Applications. ELSEVIER, 4-TH EDITION. 2013: 960.
3. Di Patre P.L., Carter D. Sternberg "S Diagnostic Surgical Pathology Review. Wolters Kluver, 2-ND Edition. 2015: 488.
4. Dietal M., Wittekind S., Bussolati G., von Winterfeld M. Pre-Analytics of Pathological Specimens In Oncology. Srringer. 2015: 133.
5. Dodd L.G, Bui M.M. ATLAS OF SOFT TISSUE AND BONE PATHOLOGY. Demosmedical. 2014: 720.
6. EDGE S.B, BYRD D.R., COMPTON C.C., FRITZ AG., GREENE F.L., TROTTI A. AJCC Cancer Staging Manual. Srringer, 7-th Edition. 2011: 646.
7. Elder D.E. LEVER "S Histopathology of the Skin. Wolters Kluver, 11-th Edition. 2014: 1544.
8. Epstein J., Netto G. Biopsy Interpretation of the Prostate. Lippincott Williams and Wilkins, 5-th Edition. 2014: 450.
9. Erickson L. Atlas of Endocrine Pathology. Srringer, 1-St Edition. 2014: 178.
10. Esmo Minimum Clinical Recommendations for Diagnosis, Treatment and Follow-Up of Cancers of Unknown Primary Site. Annals of Oncology. 2005; 16 (S.1): 175-176.
11. Fletcher c.m., Bridge J.A., Hogendoorn P., Mertens F. Who Classification of Tumours of Soft Tissue and Bone. Who Press, 4-th Edition. 2013; 5: 427.
12. KURMAN R.J., CARCANGIU M.L., HERRINGTON C.S., YOUNG R.H. WHO Classification of Tumours of the Female Reproductive Organs. Who Press, 4-th Edition. 2014; 4: 316.
13. Louis D.N., Ohgaki H., Wiestler O.D., Cavenee W.K. WHO Classification of Tumours of the Central Nervous System. Who Press, 4-th Edition. 2007; 1: 312.
14. Malpica A., E. E.D. Biopsy Interpretation of the Uterine Cervix and Corpus. Wolters Kluver, 2-ND Edition. 2015: 368.
15. Nishizuka S., Chen S.T., GWADRY F. ET AL. Diagnostic Markers That Distinguish Colon And Ovarian Adenocarcinomas: Identification by Genomic, Proteomic, and Tissue Array Profiling. Cancer Research. 2003; 63: 5243-5250.
16. Nordi Q.c. http://www.nordiqc.org.

Standardized immunohistochemical study: receptor status with breast cancer (PR, ER, KI67, HER2 NEU). It is performed only in the presence of a finished microdaparation on the slide and the tissue sample in the paraffin unit.

Russian synonyms

IHG study (RE, RP, HER2 / NEU, KI-67), immunohistochemical analysis of the receptor status of breast cancer.

Synonyms English

IHC (Immunohistochemistry) Test for Breast Cancer Receptor Status (ER, PR, HER2, KI67), Her2 Overexpression by Ihc, Estrogen Receptors, Progesterone Receptors, ER and PR Status, Estrogen and Progesterone Receptor Status.

Research method

Immunohistochemical method.

What kind of biomaterial can be used for research?

Paraffin block with bioptatom formation of breast. The primary tumor tissue can be obtained using a thick biopsy, as well as incision and excision surgical interventions. To identify metastases on a biopsy can be taken fabrics from the wall chest, regional lymph nodes or remote organs.

General research information

Modern principles and strategies for the treatment of breast cancer are based, among other things, on the results of the assessment of the receptor status and the proliferative potential of tumor cells. Tumor cells have the ability to produce and position special proteins on their surface - receptors whose stimulation leads to the launch of cellular division and tumor growth. Such receptors are able to bind to substances present in the body normally and initially in no way associated with the development of malignant neoplasms. According to the relevant clinical recommendationsFor breast cancer, the presence of the following receptors on the tumor cells of the following receptors, various combinations of which are called receptor status:

Receptors for hormones - estrogens and progesterone ( ER,Pr.). A significant part of breast tumors is hormonally dependent, that is, their growth is maintained and stimulated with estrogen and progesterone. The tumors with a positive hormonal receptor status are well aware of the therapy of hormone analogues (tamoxifen), which block the appropriate receptors - are associated with them, but do not cause activation of intracellular processes and do not allow the receptor to subsequently contact the hormone. Thus, the study of products with a tumor ER and PR makes it possible to determine its sensitivity to these drugs.

A second type receptor to the human epidermal growth factor (Human Epidermal Growth Factor Receptor 2 - HER2 / Neu.). In the cells of some breast tumors, an increased development of this receptor protein occurs, which, connecting with a natural growth factor, starts the division in the tumor cell. The total number of patients with HER2-positive breast cancer ranges from 15% to 20%. The definition of HER2 / NEU has not only prognostic value (such tumors usually progress faster and have more aggressive clinical development), but also allows you to estimate the possibility of using targeted medicinal preparations - monoclonal antibodies to the receptor HER2 - Trastuzumab (Herceptin), Lapatinib, Pertzumab. In addition, HER2-positive tumors are resistant to tamoxifen.

Proliferative activity is an indicator of the ability of tumor cells to an unlimited division, which is the main factor in the biological aggressiveness of the tumor. The division process is accompanied by the appearance of certain proteins in the cell, one of which is Ki-67.. It is not produced in cells at rest, which allows it to be used as a tumor proliferative activity marker. The determination of the level Ki-67 has an important prognostic value, since tumors from the least mature and differentiated cells have the greatest proliferative activity.

All markers mentioned above can be detected in an immunohistochemical study of a bioptate or the operating material of the tumor. To analyze from the finished paraffin unit, the finest sections are cut from a special micron, which is then attached to the glass slides and paint routine dyes to make it possible to distinguish cells from each other and from the intercellular substance. Then the cuts on the glasses are painted with solutions of antibodies labeled with fluorescent marks specific to one of the studied receptors. If the desired receptor is present in the tumor cell, the antibodies are associated with it and when viewing the glass under a special microscope, you can see fluorescence, which will indicate a positive result of the test. In addition, when viewing cut, a morphologist can see that the painted marker is located in the core, a cellular substance or on the shell of tumor cells. The amount of solutions used with antibodies corresponds to the number of markers that are investigated in the sample. The degree of fluorescence and the percentage of cells in which it is, underlie the interpretation of the results of immunohistochemical analysis and are described in more detail in the relevant section.

What is the study?

• To determine the hormonoreceptor status and the degree of proliferative activity of breast cancer for assessing the forecast and individualization of treatment, including identifying indications for the purpose of targeting therapy.
• According to the results of the detection of hormonal receptors, the feasibility of using anti-estrogen, and the receptor Her2 - targeted anti-Her2 drugs. The revealed absence of these markers avoids the appointment of obviously inefficient therapy. High index Proliferative activity, as well as negativity on receptor status for the most part, is an indication for adding cytostatic drugs to the treatment.

When is the study assigned?

• In the presence of histologically verified breast cancer - for the first time identified, recurrent and metastatic tumors.

What do the results mean?

When interpreting the results of immunohistochemical definition receptor status of steroid hormones(estrogen and progesterone) in breast tumors should be assessed not only by the percentage of cells painted by antibodies, but also the intensity of staining. Both of these parameters are recorded in the ALRED scale, where the percentage of positive cells is estimated from 0 to 5 points, and the intensity of staining from 0 to 3. The sum of the two indicators is the final score, according to which the positiveness of the tumor on receptor status is determined: 0-2 negative, 3- 8 positive. The total score 3 on this scale corresponds to 1-10% painted cells and is a minimal positive result when the purpose of hormonal therapy may have effectiveness.

Sometimes the receptor status is determined solely on the percentage of cells with painted nuclei. In such cases, NCCN recommends that all tumors are considered positive, where there are more than 1% of positive cells.

When interpreting painting on receptorHER2 /neu. Only membrane staining (staining of the cell shell), which is evaluated on a scale from 0 to +3:

the result is 0 and +1 is considered HER2-negative;

2 - a border result, with it, according to immunohistochemical research, it is impossible to judge the presence on the surface of the HER2-NEU receptor cells, it is necessary to conduct a Fish or Cish study;

3 is a positive result - anti-Her2 targeting therapy will be effective.

According to the classification of St. Gallen Consensus (2009), low proliferative activity index is considered ki-67 level less than 15%, average - 16-30%, and high - more than 30%.

What can affect the result?

• The quality of the provided paraffin blocks, the experience and qualifications of the doctors of the pathologist, since the immunohistochemical method is not fully standardized and evaluating its results to some extent subjective.
• The interpretation of the results of the study should be carried out exclusively by the doctor of the corresponding specialty, the data on the effectiveness and appropriateness of the appointment of certain medicines Depending on the results of the study, we are exceptionally advisory and may be revised taking into account the individual characteristics of the patient.

Important comments

• With an indefinite Her2 / Neu-receptor status (the result of immunohistochemical research 2+), the execution of Fish or Cish-study is recommended that will eliminate the hyperactivation of the gene encoding this receptor. With the unavailability of these studies, a repeated immunohistochemical study is allowed, but on another sample of tumor tissue.
• There are several scales for estimating the receptor status of breast cancer, in the laboratory report should be indicated, for which the assessment of the positiveness of the tumor in this study was carried out, and the descriptive characteristic of the number of positive cells, the features of staining cell structures and the morphological features of the cells was given.

Histological study of biopsy breast biopsy

Cytological study of breast points

Determination of HER2 Tumor Status by Fish

Determination of HER2 Tumor Status by CISH

Who appoints a study?

Oncologist, mammologist, onkogynecologist.

Literature

Dana Carmen Zaha. Significance of ImmunoHistochemistry in Breast Cancer. World journal of clinical oncology, 2014; 5 (3): 382-392.

NCCN CLINICAL PRACTICE GUIDELINES IN ONCOLOGY: BREAST CANCER. Version 3.2017 - November 10, 2017. Available at www.nccn.org.

Clinical laboratory diagnostics: National Guide: In 2 T. - T. I / Ed. V. V. Dolgova, V. V. Menshikova. - M.: Goeotar Media, 2012. P. 658-660.

V. F. Semigzov, R. M. Kaltuyev, V. V. Semilzov, G. A. Dashyan, T. Yu. Semilazova, P. V. Krivorownko, K. S. Nikolaev. General recommendations on the treatment of early breast cancer St. GALLE-2015, adapted by the experts of the Russian society oncommologists. Tumors of the female reproductive system, 2015; 3: 43-60.

Alternative names: IGH, English: Immunohistochemistry or IHC.

Immunohistochemical study is a special method of diagnosing tumor diseases. The essence of the method is to study under the microscope images of tissues previously treated with specific antibodies.

Currently, antibodies have been created practically to all tumors.

With the help of immunohistochemical research:

Indications for immunohistochemical research

With this method, you can explore any fabrics. The main indication to carry out a suspicion of the tumor process.

The following readings are defined for IGI:

Contraindications

TO this method Research no contraindications. Its impossible is impossible only if there is no possibility to obtain biopsy material.

How is an immunohistochemical study

The study itself consists of four stages:

The results of the study become known after 7 days (with a standard study - "small panel") or through 15 (an extended study is a "large panel").

Interpretation of results

The study of samples is engaged in a pathologist who has passed special training in IGG. In conclusion, the doctor notes to which antibodies the tropiness (affinity) of the tissue is determined. Additionally, the morphological structure of the sample is described - which cells and in what quantity are present.

Identification of tissue affinity to certain standard tumor antibodies indicates a specific form of an oncological disease.

Additional Information

Immunohistochemical study is currently the most accurate method for diagnosing tumor diseases. It allows you to put the final diagnosis with an accuracy of 99%, determine the type of tumor, reveal its primary localization.

Literature: