Purpose. The histological diagnosis of Mycobacterium tuberculosis (MTB) remains a diagnostic challenge despite different methods. Immunohistochemistry (IHC) not only could confirm granulomatous tissue involvement but also can demonstrate MTB antigen immunolocalization. This study tries to clarify the details of immunohistochemical staining for MTB with pAbBCG. Materials/Methods. Twenty-three confirmed TB granulomatous tissue samples were studied by Ziehl-Neelsen and immunohistochemistry (IHC) staining with pAbBCG. Samples were selected from the archive of the Department of Pathology, National Research Institute of Tuberculosis and Lung Disease, Tehran, Iran. Results. IHC staining was positive in all samples, whereas Ziehl-Neelsen was positive in 9 cases out of 23 (39.1%). Tissue types used were pleural tissue, lymph nodes, and lung tissue. IHC showed positive coarse granular cytoplasmic and round, fragmented bacillary staining. In this study, epithelioid cells clearly showed more positive staining at the periphery of the granuloma rather than the center of granuloma. There is also positive staining in endothelial cells, fibroblasts, plasma cells, lymphocytes, and macrophages outside the granuloma. Conclusion. Considering the criteria of positive immunohistochemical staining of TB granulomatous reactions, this stain not only highlights the presence of mycobacterial antigens for tissue diagnosis, but also could morphologically localize its distribution in different cells. 1. Introduction Histological diagnosis of tuberculosis (TB) has long been an important issue in anatomical pathology. On the other hand, extrapulmonary TB comprises 10–15% of infections with Mtb. Following HIV prevalence increase, extrapulmonary TB is rising as well [1, 2]. Considering the limitations in sensitivity and specificity of Ziehl-Neelsen staining [3], mycobacterial evaluation, mycobacterial culture, and molecular and serological techniques [4–6], histomorphological analysis appears to be the only feasible technique for field diagnosis of TB in some patients [7, 8]. Granulomatous reactions, and in some cases, nongranulomatous reactions such as the presence of foamy macrophages [9] or mycobacterial spindle cell pseudotumour [10] in some types of mycobacterial infections are indicative of TB only if the presence of TB bacilli has been confirmed in the tissue. Recently, it has been stated that PCR results are acceptable only if the presence of TB bacilli can be confirmed in the tissue [11]. Detection of beaded bacilli (which are more commonly observed in the necrotic zone) by
References
[1]
G. T. Programme, Global Tuberculosis Control: WHO Report: Global Tuberculosis Programme, World Health Organization, 2010.
[2]
E. L. Corbett, C. J. Watt, N. Walker et al., “The growing burden of tuberculosis: global trends and interactions with the HIV epidemic,” Archives of Internal Medicine, vol. 163, no. 9, pp. 1009–1021, 2003.
[3]
A. Trusov, R. Bumgarner, R. Valijev et al., “Comparison of Lumin？ LED fluorescent attachment, fluorescent microscopy and Ziehl-Neelsen for AFB diagnosis,” International Journal of Tuberculosis and Lung Disease, vol. 13, no. 7, pp. 836–841, 2009.
[4]
A. M. Attallah, C. A. A. Malak, H. Ismail, A. H. El-Saggan, M. M. Omran, and A. A. Tabll, “Rapid and simple detection of a Mycobacterium tuberculosis circulating antigen in serum using dot-ELISA for field diagnosis of pulmonary tuberculosis,” Journal of Immunoassay and Immunochemistry, vol. 24, no. 1, pp. 73–87, 2003.
[5]
J. M. Dora, G. Geib, R. Chakr et al., “Polymerase chain reaction as a useful and simple tool for rapid diagnosis of tuberculous meningitis in a Brazilian tertiary care hospital,” Brazilian Journal of Infectious Diseases, vol. 12, no. 3, pp. 245–247, 2008.
[6]
A. Mishra, A. Singhal, D. S. Chauhan et al., “Direct detection and identification of Mycobacterium tuberculosis and Mycobacterium bovis in bovine samples by a novel nested PCR assay: correlation with conventional techniques,” Journal of Clinical Microbiology, vol. 43, no. 11, pp. 5670–5678, 2005.
[7]
J. S. Pedersen, I. Clarke, and J. Mills, “Improved detection of mycobacteria species in formalin-fixed tissue sections,” Histopathology, vol. 59, no. 5, pp. 993–1005, 2011.
[8]
T. Mustafa, H. G. Wiker, S. G. M. Mfinanga, O. M？rkve, and L. Sviland, “Immunohistochemistry using a Mycobacterium tuberculosis complex specific antibody for improved diagnosis of tuberculous lymphadenitis,” Modern Pathology, vol. 19, no. 12, pp. 1606–1614, 2006.
[9]
H. D'Avila, C. M. Maya-Monteiro, and P. T. Bozza, “Lipid bodies in innate immune response to bacterial and parasite infections,” International Immunopharmacology, vol. 8, no. 10, pp. 1308–1315, 2008.
[10]
S. Logani, D. R. Lucas, J. D. Cheng, H. L. Ioachim, and N. V. Adsay, “Spindle cell tumors associated with mycobacteria in lymph nodes of HIV-positive patients: ‘Kaposi sarcoma with mycobacteria’ and ‘mycobacterial pseudotumor’,” American Journal of Surgical Pathology, vol. 23, no. 6, pp. 656–661, 1999.
[11]
T. Ulrichs, M. Lefmann, M. Reich et al., “Modified immunohistological staining allows detection of Ziehl-Neelsen-negative Mycobacterium tuberculosis organisms and their precise localization in human tissue,” Journal of Pathology, vol. 205, no. 5, pp. 633–640, 2005.
[12]
á. Somosk？vi, J. E. Hotaling, M. Fitzgerald, D. O'Donnell, L. M. Parsons, and M. Salfinger, “Lessons from a proficiency testing event for acid-fast microscopy,” Chest, vol. 120, no. 1, pp. 250–257, 2001.
[13]
R. W. Smithwìck, Laboratory Manual For Acid-Fast Microscopy: Public Health Service, Center For Disease Control, Bureau of Laboratories, 1976.
[14]
M. M. Goel and P. Budhwar, “Immunohistochemical localization of mycobacterium tuberculosis complex antigen with antibody to 38 kDa antigen versus Ziehl Neelsen staining in tissue granulomas of extrapulmonary tuberculosis,” The Indian journal of tuberculosis, vol. 54, no. 1, pp. 24–29, 2007.
[15]
M. R. Purohit, T. Mustafa, H. G. Wiker, O. M？rkve, and L. Sviland, “Immunohistochemical diagnosis of abdominal and lymph node tuberculosis by detecting Mycobacterium tuberculosis complex specific antigen MPT64,” Diagnostic Pathology, vol. 2, no. 1, article 36, 2007.
[16]
A. A. Velayati, P. Farnia, M. R. Masjedi, and G. Konstantinovich, “Morphological modification by tubercle bacilli: no time for denial,” Journal of Infection in Developing Countries, vol. 6, no. 1, pp. 97–99, 2012.
[17]
K. M. Dobos, E. A. Spotts, F. D. Quinn, and C. H. King, “Necrosis of lung epithelial cells during infection with Mycobacterium tuberculosis is preceded by cell permeation,” Infection and Immunity, vol. 68, no. 11, pp. 6300–6310, 2000.
[18]
S. A. Oracki, J. A. Walker, M. L. Hibbs, L. M. Corcoran, and D. M. Tarlinton, “Plasma cell development and survival,” Immunological Reviews, vol. 237, no. 1, pp. 140–159, 2010.
[19]
G. González-Avila, C. Sandoval, M. T. Herrera et al., “Mycobacterium tuberculosis effects on fibroblast collagen metabolism,” Respiration, vol. 77, no. 2, pp. 195–202, 2009.
