Indian Journal of Dermatology
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BASIC RESEARCH
Year : 2007  |  Volume : 52  |  Issue : 3  |  Page : 131-133
Pan B and T cell markers in cutaneous tuberculosis: An immunohistochemical study


1 From the Urban Health Center, Division of Community Medicine, India
2 Rajah Muthiah Medical College, Annamalai University, Annamalai Nagar, Chidambaram - 608002, TN, India
3 Tuberculosis Research Center, Chetput, Chennai, India

Correspondence Address:
L Padmavathy
B3, RSA Complex, Annamalai University, Annamalai Nagar - 608002, Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0019-5154.35090

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   Abstract 

Histopathology of tuberculosis and leprosy is expected to correlate with the clinical and gross pathological aspects of the disease. However, this may not be the case, particularly in tuberculosis. This reflects our inadequate interpretation of the cellular events in tissues. The advent of immunocytochemistry for finding the cell types in lesions and some of their functions have helped in the interpretation of these changes. Immunocytochemical studies using monoclonal antibodies have shown consistent relationship between histology and lymphocyte organization in leprosy, a related mycobacterial disease. The study of the patterns of cellular responses in the granuloma per se from the different clinical expressions of tuberculosis is essential to comprehend the different immunopathogenic mechanisms in each. CD stands for cluster designation and is a nomenclature for the identification of specific cell surface antigens defined as monoclonal antibodies. This procedure can be applied to both formalin-fixed and frozen tissues. It is very helpful in identifying subpopulations of T and B cell lymphocytes. CD3 T cell is marked by receptors for antigen recognition, while CD20 is meant for Pan B cells and dendritic cells. Since CMI is the preeminent immune response to the Mycobacterium tuberculosis infection, interaction of antigen-presenting cells with antigen specific B and T lymphocytes is of paramount importance, which should be accompanied by cytokine production and liberation. The present study is undertaken for evaluating the cell types in lesions of cutaneous tuberculosis and some of their functions using B and T cell markers.


Keywords: CD20 cells, CD3, immuno-histochemistry, monoclonal antibodies, T and B cells


How to cite this article:
Padmavathy L, Rao LL, Shakila H, Ramanathan V D. Pan B and T cell markers in cutaneous tuberculosis: An immunohistochemical study. Indian J Dermatol 2007;52:131-3

How to cite this URL:
Padmavathy L, Rao LL, Shakila H, Ramanathan V D. Pan B and T cell markers in cutaneous tuberculosis: An immunohistochemical study. Indian J Dermatol [serial online] 2007 [cited 2019 Nov 22];52:131-3. Available from: http://www.e-ijd.org/text.asp?2007/52/3/131/35090



   Introduction Top


Tuberculosis still remains a scourge affecting the developing nations inspite of the measures introduced to reduce the incidence. Extrapulmonary tuberculosis affecting other systems such as CNS, bones and joints and skin is also common in these areas. The demonstration of Mycobacterium tuberculosis, which is essential for confirming the diagnosis, is difficult and time consuming in cases of extrapulmonary tuberculosis, particularly in cutaneous tuberculosis as the bacillary load is considerably low. The study of the patterns of cellular responses in the granuloma per se from the different clinical expressions of the disease is essential to comprehend the different immunopathogenic mechanisms in each. [1] The B and T cell marker analysis would be of use in such studies.


   Materials and Methods Top


One hundred and seventeen patients who attended the Dermatology Department of Rajah Muthiah Medical College and Hospital, Annamalai University, with various clinical types of cutaneous tuberculosis between 1991 and 1998 were included in this series. The clinical diagnosis of cutaneous tuberculosis was corroborated by biopsy and histopathological examination.

Immunohistochemical study was undertaken to demonstrate T and B cell populations in the tissue sections in the selected cases of cutaneous tuberculosis.

a) By the immunoperoxidase staining method, paraffin sections from 52 biopsy specimens were subjected to CD20 (B cells) marker studies.

b) Paraffin sections from 41 biopsies were subjected to CD3 (T cell) marker studies.


   B cell (CD20) and T cell (CD3) staining Top


In order to find out the type of cells present in the histological sections, anti-CD20 staining and anti-CD3 staining were performed to locate B cells and T cells, respectively. Immunoenzymatic staining was done on paraffin sections using standard indirect immunoperoxidase staining (peroxidase-antiperoxidase staining).

Principle: The principle is similar to that of the enzyme-linked immunosorbent assay. The tissue containing the antigen is treated with an appropriate antibody, to form an antigen-antibody complex. The section is again treated with peroxidase conjugated secondary antibody, and the antigen-antibody complex binds with the conjugate. When the substrate is added to this, the brown colored areas indicate the occurrence of the reaction. These brown colored areas are the positive sites of the location of antigen-antibody complexes. [2],[3]

Antiserum used: Anti H37 RV serum raised in rabbits.


   Observations Top


The present study is a prospective study in which random biopsy specimens were subjected to T- and B-cell-marker studies.

Among the 52 biopsy specimens subjected to CD20 (B cells) marker studies, an overall positivity of 10% was noted for CD20, the highest being among scrofuloderma (16%), followed by LV (7.6%) and TBVC (6.5%). The positive cells were observed predominantly within the granuloma [Figure - 1]. In these cases, the caseation necrosis was prominent, particularly in cases of scrofuloderma.

From the 41 biopsy sections subjected to CD3 (T cells) marker studies, an overall positivity of 28.3% was recorded for CD3, with Lupus vulgaris showing the highest positivity (35%), followed by TBVC (30%) and scrofuloderma (20%). The CD3 positive cells were seen predominantly within the granuloma [Figure - 2].

In the 36 skin biopsies in which both T and B cell marker studies were undertaken, T cells were predominantly observed along with a few B cells in the granulomas in all the cases. B cells were relatively more in scrofuloderma than in LV and TBVC.

The response to treatment was better among cases in which T cells were predominant.


   Discussion Top


It is reasonable to expect that histopathology should correlate with the clinical and gross pathological aspects of tuberculosis and leprosy. However, it may not, particularly in the case of tuberculosis, and this reflects our insufficient interpretation of the cellular events in tissues. [4] The advent of immunocytochemistry for evaluating the cell types in lesions and some of their functions has helped in the interpretation of these changes.

Immunocytochemical studies using monoclonal antibodies have shown consistent relationship between histology and lymphocyte organization in leprosy, a related mycobacterial disease. [4]

B cells: An overall positivity of 10% was recorded for CD-20 cells, the value being highest in scrofuloderma. The number of B cells was higher in scrofuloderma (16%), but it was comparable in TBVC (6.5%) and LV (7.6%). Thus, the B cells were more often associated with necrosis as in scrofuloderma. In a study conducted in Chennai, 23% B cells in scrofuloderma, 12.5% in TBVC and 10% in lupus vulgaris were reported. [3] A similar increase in B cell numbers was documented in the cases of scrofuloderma, where the anti-CD22 antibody is used to stain the B cells in cases of cutaneous tuberculosis. [1]

CD3 cells : The number of CD3 positive cells was comparable in lupus vulgaris (35%) and TBVC (30%), while they were significantly less in scrofuloderma (20%).

In a study involving 120 cases reported from Chennai, CD3 cells were observed up to 75% of the total lymphocytes in LV, 81% in TBVC and 63% in scrofuloderma. [5] Similar findings using CD11 antibodies were reported in other studies. [1]

The plasma cells generated from the B cells can produce antibodies and can form in situ immune complexes along with the fragmented M. tuberculosis antigen. [6] This phenomenon can not only initiate but also determine the type of granuloma formed. [7],[8] It was shown that anti-idiotypic B cells against M. tuberculosis can modulate both granuloma formation and a delayed hypersensitivity reaction to PPD. [9] Antigen-specific B lymphocytes are potent antigen-presenting cells. It is known that if B cells present antigen to the naοve T lymphocytes, it results in tolerance, whereas if they present antigen to sensitized T cells, they can be immunized. [10]

The presence of antigen, plasma cells and T lymphocytes in the lesions together suggests that the in situ formation of antigen-antibody complexes is possibly responsible for the necrosis observed in scrofuloderma. The role of immune complexes in the production of some of the other features of the granuloma is also well recognized.

Considering that CMI is the preeminent immune response to the M. tuberculosis infection, the interaction of antigen-presenting cells with antigen specific B and T lymphocytes is of paramount importance, which should be accompanied by cytokine production and liberation. [1] This is an intricate process, and many facets of this process are still unaccounted for or poorly documented. [11],[12] It was suggested that tuberculosis can be classified into two polar groups based on the cellular response: the first group showing good cell-mediated immunity and very small or no antibody formation and the second group showing poor cellular responses and excessive antibody production. [13]


   Acknowledgement Top


We would like to thank the authorities for the facilities provided for carrying out the work and for permission to publish this article.

 
   References Top

1.Sehgal VN, Gupta R, Bose M, Saha K. Immunohistopathological spectrum in cutaneous tuberculosis. Clin Exp Dermatol 1993;18:309.  Back to cited text no. 1      
2.Orell JM, Brett SJ, Ivanyi J, Coghill G, Grant A, Swansonbeck J. Measurement of tissue distribution of immunoperoxidase staining with polyclonal anti-BCG serum in lung granulomata of mice infected with Mycobacterium tuberculosis . J Pathol 1991;164:41.  Back to cited text no. 2      
3.Graham RC, Karnovsky J. The early stages of absorption of injected horse-radish peroxidase in the proximal tubules of mouse kidney. Ultra structural cytochemistry by a new technology. J Histochem Cytochem 1966;14:21.  Back to cited text no. 3      
4.Lucas SB. Histopathology of leprosy and tuberculosis: An overview. Br Med Bull 1988;44:584-99.  Back to cited text no. 4      
5.Shakila H. Immunopathological studies of naturally occurring and experimental dermal granuloma induced by Mycobacterium tuberculosis. Thesis submitted to The Tamil Nadu Dr. M. G. R. Medical University, March 1977.  Back to cited text no. 5      
6.Ramanathan VD, Shakila H, Umapathy KC. The induction of a 'Mitsuda' type response by purified protein derivative (PPD) in tuberculosis. Proceedings of the 10 th International Congress of Immunology. New Delhi; 1998.  Back to cited text no. 6      
7.Spector WG, Heesom N. The production of granulomata by antigen-antibody complexes. J Pathol 1969;98:31.  Back to cited text no. 7      
8.Ridley MJ, Marianayagam Y, Spector WG. Experimental granulomas induced by mycobacterial immune complexes. J Pathol 1982;136:59-72.  Back to cited text no. 8      
9.Campa M, Marelli P, Ota F. B cell mediated depression of granulomatous response to BCG in mice. Cell Immunol 1989;119:279-85.  Back to cited text no. 9      
10.Matzinger P. Tolerance, danger and extended family. Ann Rev Immunol 1994;12:991-1045.  Back to cited text no. 10      
11.Boom WH, Wallis RS, Chervena KA. Mycobacterium tuberculosis reactive CD+T cell clones: Heterogeneity in antigen recognition, cytokine production and cyotoxicity for mononuclear phagocytes. Infect Immun 1991;59:2737-43.  Back to cited text no. 11      
12.Shivatsuchi H, Johnson JL, Ellner J. Bidirectional effects of cytokines on the growth of Mycobacterium avium within human monocytes. J Immunol 1991;146:3165-70.  Back to cited text no. 12      
13.Lenzini L, Rottoli P, Rottoli L. The spectrum of human tuberculosis. Clin Exp Immunol 1977;27:230-7.  Back to cited text no. 13      


    Figures

  [Figure - 1], [Figure - 2]



 

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    Abstract
    Introduction
    Materials and Me...
    B cell (CD20) an...
    Observations
    Discussion
    Acknowledgement
    References
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