Comparative characteristics of immunohistochemical detection of M. tuberculosis antigens and mycobacteria by Ziehl–Neelsen method in lung tissue with tuberculoma during progressive course of tuberculosis

Authors

  • I. V. Liskina SI “National Institute of Phthisiology and Pulmonology named after F. G. Yanovsky of NAMS of Ukraine”, Kyiv,
  • O. A. Melnik SI “National Institute of Phthisiology and Pulmonology named after F. G. Yanovsky of NAMS of Ukraine”, Kyiv,
  • L. M. Zahaba SI “National Institute of Phthisiology and Pulmonology named after F. G. Yanovsky of NAMS of Ukraine”, Kyiv,
  • S. D. Kuzovkova SI “National Institute of Phthisiology and Pulmonology named after F. G. Yanovsky of NAMS of Ukraine”, Kyiv,
  • V. V. Kuts SI “National Institute of Phthisiology and Pulmonology named after F. G. Yanovsky of NAMS of Ukraine”, Kyiv,

DOI:

https://doi.org/10.14739/2310-1237.2017.3.118754

Keywords:

pulmonary tuberculoma, M. tuberculosis, antigens, immunohistochemistry, histocytochemistry

Abstract

The aim of the study – to compare the character of expression of M. tuberculosis antigens in immunohistochemical (IHC) study and detection of acid-fast bacilli (AFB) by Ziehl–Neelsen method in lung tissue with tuberculoma in the chronic course of tuberculosis during the exacerbation phase.

Materials and methods. The materials for the study were resected lungs, in total 19 cases. The thickness of the serial sections was 5–6 microns. Staining was carried out with hematoxylin and eosin, by Ziehl–Neelsen, and the IHC.

IHC study used a rabbit polyclonal antibody to Mycobacterium tuberculosis. Antibody (USA) was used at 1:2500 dilution. Assessing of the positive reaction was according to the conditional scale: + – background color, pale yellow, diffuse; ++ – fine-granular (light brown); +++ – coarse-granular (intense brown). Quantitative gradation of AFB was the following: out or within macrophages in one field of view: few, 1–5 units; a moderate amount – 6–15, and many – more than 15.

Results. In all the cases pulmonary tuberculosis had morphological signs of the progression of a specific inflammatory process.

Ziehl–Neelsen stain showed that the most frequently free located AFB were detected in the necrotic nucleus (84.2 %) and granulation layer of the capsule of tuberculoma (68.4 %). Most often single bacilli were noted. A greater number of bacilli were found inside macrophages.

The most frequently intracellular bacilli were found in macrophages located in alveolar spaces – 84.2 % observations in a moderate number.

IHC study showed that in all the cases, cells with a coarse-granular positivity (+++) to MBT antigens were determined. Macrophages within the alveoli outside tuberculoma, with same positivity were in 94.7 %; fewer – macrophages and epithelioid cells were in the granulation layer of the tuberculoma capsule, a total of 84.2 %. The percentage of observations of a large number of cells in the alveoli reached 77.8 %, and in the granulation layer of the capsule – 30.0–50.0 %.

More often the fine-granular staining of cells in the same parts of lung tissue was observed, a cases of large and/or moderate number of cells prevailed.

Conclusions. The AFB was identified in 94.7 % cases by Ziehl–Neelsen method. In IHC study a positive reaction of macrophage cells was detected in all the cases (100 %).

IHC study showed that maximum antigenic load was determined in macrophages localized inside alveoli outside of tuberculoma and slightly less in macrophage cells of the granulation layer of the tuberculoma’s capsule. The results of both methods for detecting mycobacteria and/or their fragments demonstrated the coincidence of the maximum number of observations of localization of the bacilli structures, namely in granulation layer of the tuberculoma’s capsule and the preserved alveolar spaces outside the tuberculoma.

References

Averbakh, M. M. (1962). Tuberkulomy legkogo. [Pulmonary tuberculoma]. Moscow: Gos. izd-vo med. literatury [in Russian].

DU «Ukrainskyi tsentr kontroliu za sotsialno nebezpechnymy khvorobamy Ministerstva okhorony zdorov’ia Ukrainy» (2016) VIL-infektsiia v Ukraini. [HIV-infection in Ukraine]. Informatsiinyi biuleten, 46. [in Ukrainian].

Liskina, I. V., Kuzovkova, S. D., Kravchenko, S. O., Zahaba, L. M., & Lukianchuk, V. H. (2010). Histolohichna diahnostyka stupenia aktyvnosti tuberkuloznoho zapalnoho protsesu pry tuberkulomakh lehen [Histological diagnostics of the activity degree of tuberculosis inflammatory process in pulmonary tuberculoma]. Kyiv. [in Ukrainian].

Rekalova, O. M., Belohortseva, O. I., & Koval, N. G. (2017). Imunolohichni metody diahnostyky tuberkulozu [Immunological methods of diagnosis of tuberculosis]. Tuberkuloz, lehenevi khvoroby, VIL-infektsiia, 1, 75–81. [in Ukrainian].

Siev, M., Wilson, D., Kainth, S., Kasprowicz, V. O., Feintuch, C. M., Jenny-Avital, E. R., & Achkar, J. M. (2014) Antibodies against mycobacterial proteins as biomarkers for HIV-associated smear-negative tuberculosis. Clin. Vaccine Immunol, 21(6):791-8. doi: 10.1128/CVI.00805-13.

Trusov, A., Bumgarner, R., Valijev, R., Chestnova, R., Talevski, S., Vragoterova, C., & Neeley, E. S. (2009) Comparison of LuminŮ LED fluorescent attachment, fluorescent microscopy and Ziehl-Neelsen for AFB diagnosis. International Journal of Tuberculosis and Lung Disease, 13(7), 836–41.

D'Avila, H., Maya-Monteiro, C. M., & Bozza, P. T. (2008) Lipid bodies in innate immune response to bacterial and parasite infections. International Immunopharmacology, 8(10):1308-15. doi: 10.1016/j.intimp.2008.01.035

Mustafa, T., Wiker, H. G., Mfinanga, S. G., Mørkve, O., & Sviland, L. (2006). Immunohistochemistry using a Mycobacterium tuberculosis complex specific antibody for improved diagnosis of tuberculous lymphadenitis. Modern Pathology, 9(12), 1606–14. doi: 10.1038/modpathol.3800697

Purohit, M. R., Mustafa, T., Wiker, H. G., Mørkve, O., & Sviland, L. (2007). Immunohistochemical diagnosis of abdominal and lymph node tuberculosis by detecting Mycobacterium tuberculosis complex specific antigen MPT64. Diagnostic Pathology, 2, 36. doi: 10.1186/1746-1596-2-36

World Health Organization (2010) Global Tuberculosis Control: WHO Report. Geneva. Retrieved from http://reliefweb.int/sites/reliefweb.int/files/resources/F530290AD0279399C12577D8003E9D65-Full_Report.pdf.

Guirado, E., Schlesinger, L. S., & Kaplan, G. (2013). Macrophages in tuberculosis: friend or foe. Semin. Immunopathol, 35(5), 563–83. doi: 10.1007/s00281-013-0388-2.

Karimi, S., Shamaei, M., Pourabdollah, M., Sadr, M., Karbasi, M., Kiani, A., & Bahadori, M. (2014). Histopathological findings in immunohistological staining of the granulomatous tissue reaction associated with tuberculosis. Tuberculosis Research and Treatment, 2014. doi: 10.1155/2014/858396.

Lawn, S. D., & Zumla, A. I. (2011). Tuberculosis. Lancet, 378, 57–72.

Somoskövi, A., Hotaling, J. E., Fitzgerald, M., O'Donnell, D., Parsons, L. M., & Salfinger, M. (2001). Lessons from a proficiency testing event for acid-fast microscopy. Chest, 120(1), 250–257. doi: 10.1378/chest.120.1.250.

Ulrichs, T., Lefmann, M., Reich, M., Morawietz, L., Roth, A., Brinkmann, V., et al. (2005). Modified immunohistological staining allows detection of Ziehl–Neelsen-negative Mycobacterium tuberculosis organisms and their precise localization in human tissue. Journal Pathology, 205, 633–640. doi: 10.1002/path.1728

Pedersen, J. S., Clarke, I., & Mills, J. (2011). Improved detection of mycobacteria species in formalin-fixed tissue sections. Histopathology, 59(5), 993–1005. doi: 10.1111/j.1365-2559.2011.04015.x.

Ryu, Y. J. (2015). Diagnosis of pulmonary tuberculosis: recent advances and diagnostic algorithms. Tuberculosis Respiratory Disease, 78, 64–71. doi: 10.4046/trd.2015.78.2.64.

Logani, S., Lucas, D. R., Cheng, J. D., Ioachim, H. L., & Adsay, N.V. (1999). 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, 23(6), 656–661.

Corbett, E. L., Watt, C. J., Walker, N., Maher, D., Williams, B. G., Raviglione, M. C., & Dye, C. (2003). The growing burden of tuberculosis: global trends and interactions with the HIV epidemic. Archives of Internal Medicine, 163(9), 1009–1021. doi: 10.1001/archinte.163.9.1009.

Downloads

How to Cite

1.
Liskina IV, Melnik OA, Zahaba LM, Kuzovkova SD, Kuts VV. Comparative characteristics of immunohistochemical detection of M. tuberculosis antigens and mycobacteria by Ziehl–Neelsen method in lung tissue with tuberculoma during progressive course of tuberculosis. Pathologia [Internet]. 2017Dec.22 [cited 2024Nov.13];(3). Available from: http://pat.zsmu.edu.ua/article/view/118754

Issue

No. 3 (2017)

Section

Original research

License

Authors who publish with this journal agree to the following terms:

    1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
Лицензия Creative Commons
    1. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.

  1. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (SeeThe Effect of Open Access).