To Analyse the Differentiation and Grading of Non-Hodgkins Lymphoma by Immunohistochemical Expression of CD20, CD3 and Ki67 in Lymph Node

Immunophenotyping, although has emerged indispensable in the diagnosis and classification of lymphoid neoplasms, has to be used cautiously with knowledge of the antibodies used. No antigen is totally lineage or lymphoma specific, and for this reason, immunostaining must be performed in the context of a panel. In addition, familiarity with the diagnostic criteria and differential diagnosis of each lymphoid tumor and ultimately correlation with morphology, and clinical history are essential to enhance the diagnostic accuracy and reproducibility. AIM: The present retrospective study aimed to analyse the differentiation and grading of Non-Hodgkin’s lymphoma by immunohistochemical expression of CD20, CD3 and Ki-67 in lymph node. Methods: A total of 50 samples of NHL were included in the study. Written informed consent of the patient was taken where ever required in the vernacular. Relevant history of the patient was also taken as per the proforma attached along with. The tissues were stained with H and E staining, CD20, CD 3 and Ki-67 immunostaining. The positive immunostained slides were then evaluated and scored both qualitatively and quantitatively. Results: In the present study a total of 50 samples of NHL were included with an age range from 12 to 72 years and a mean age of 46.54 years and male predominance.45 cases showed immunopositivity for CD 20, showing that they belong to B cell phenotype and only 5 cases showed immunopositivity for CD 3, thus showing T cell phenotype. The mean Ki-67 for B cell lymphoma patients was 47.86 ±28.04,with a minimum score of 2 and a max score of 92 and for T cell lymphoma patients was 61.4±18.02, with a minimum score of 40 and a max score of 81, but there was no significant correlation between them (P=0.382). Conclusion: Ki-67 expression in NHL can help in monitoring of patients at risk and can to some extent also aid in detecting the degree of aggressiveness of the disease to give suitable treatment but Ki-67 alone cannot be a risk factor in NHL patients and other factors such as age, sex and type of NHL can be affective, too. The outcome of further analyzing the association between Ki-67 expression and the prognosis of various subtypes of lymphoma should be supported.

Diagnostic Utility of Cell Block of Bronchial Brush Tip Washing in the Diagnosis of Carcinoma Lung and Application of Cytokeratin 5/6, TTF-1 & CD 56 Expression for Subclassification

Lung cancer is one of the most common malignancies worldwide with high mortality. Furthermore being one of the most insidious and aggressive neoplasm in the realm of oncology, it’s timely diagnosis and accurate subclassification becomes pre-requisite for administering appropriate and timely target therapy. In the present study, cell block from brush tip washings were prepared and immunoreactivity assessed for cytokeratin5/6, TTF-1 and CD56 with aim to diagnose and sub-classify carcinoma lung. Methods: The present study was conducted on 25 specimens of brushtip washings from suspected cases of carcinoma lung. Bronchoscopic investigation of pulmonary lesions was performed and routine brush smears were made and these brush tip were processed into cell block. Immunohistochemical staining for marker CK5/6, CD56 & TTF 1 was done and evaluated. Results: Brush smear cytology finding was mostly benign seen in 12 (48%) followed by SCC seen in 4 (16%), ADC in 3 (12%). Cell block microscopy showed SCC in 11 (44%), ADC in 5 (20%), small cell carcinoma in 3 (12%) and carcinoma in 3 (12%). It was inadequate in 3 (12%). The difference was significant (P< 0.05). Sensitivity and specificity of brush smear cytology in diagnosing lesion was 33.3 % and 52.6% respectively.Whereas for cell block microscopy in diagnosing lesion sensitivity was 91.67% and specificity 86.6%. The overall sensitivity of IHC CK 5/6 in diagnosing SCC was 100% and specificity was 52.4%. CD56, TTF1 were negative in these cases. CD56 showed 100% sensitivity in diagnosing small cell carcinoma with specificity being 24.6%, The overall sensitivity of IHC TTF 1 in diagnosing ADC was 100% & for small cell carcinoma was 40%. Conclusion: Cell block preparation is a simple method that increases diagnostic yield of flexible bronchoscopy, is cost effective & hence can be routinely used. IHC panel consisting CK 5/6, CD 56 and TTF 1 has more diagnostic value in precise subtyping of different types of lung carcinoma in adjunction to routine H&E staining.

Role of Adenosine Deaminase in Pleural Effusions and its Cytological Correlation

Diagnosis of pleural diseases creates difficulty due to overlapping features of various benign and malignant conditions. However pleural fluid cytology of closed pleural biopsy specimen is most commonly used procedure to diagnose various pleural diseases in developing country like India. Present study was conducted to establish diagnostic utility of ADA in Pleural Fluids and its correlation with cytological findings. Methods: The present study includes 100 samples of pleural fluid samples taken from patients coming to the Department of Chest and TB, Government Medical College and Hospital, Amritsar with the complaint of pleural effusion. Results: For ADA levels in pleural fluid Sensitivity is 92%, Specificity is 81.33%, Positive predictive value is 62.16% and Negative predictive value is 96.83%. For lymphocyte count in pleural fluid Sensitivity is 100%, Specificity is 16.67%, Positive predictive value is 31.25 % and Negative predictive value is 100%. In Combination of ADA and lymphocyte count to diagnose tubercular pleural effusion Sensitivity is 100%, Specificity is 77.27%, Positive predictive value is 82.14 % and Negative predictive value is 100%. Conclusion: Measurement of ADA level in pleural fluid in combination with the differential count of pleural fluid will give best results to categorize and to rule in the diagnosis of tubercular pleural effusion.

Immunological unresponsiveness in leprosy and its relevance to immunoregulation in man.

The varied forms of leprosy form a clinical and immunological spectrum which offers extraordinary possibilities for insight into immunoregulatory mechanisms in man. At one pole, tuberculoid leprosy, patients develop high levels of cell-mediated immunity which ultimately results in killing of bacilli in the tissues, albeit often with damage to nerves. At the lepromatous pole, patients exhibit selective immunological unresponsiveness to antigens of Mycobacterium leprae. Even though all currently known protein species of Mycobacterium leprae and BCG are cross-reactive, lepromatous patients unreactive to Mycobacterium leprae antigens frequently respond strongly to tuberculin. In vitro experiments suggest the existence of lepromin-induced suppressor activity, mediated by both monocytes and Τ cells. The Τ suppressor cells have the T8 phenotype of which 50% express the activation markers, Ia and FcR. The one unique species of antigen of the leprosy bacillus is a phenolic glycolipid, and it appears that the Ts cells largely recognize the terminal trisaccharide of this unique antigen. Depletion of Ts cells restores in vitro reactivity of lymphocytes to lepromin in a portion of lepromatous patients, and addition of IL-2 containing supernatants partially restores responsiveness to Mycobacterium leprae antigens. Vaccination of lepromatous patients with a mixture of Mycobacterium leprae and live BCG restores cell-mediated immunity in the majority of lepromatous patients, and concomitantly reduces the in vitro suppressor activity and number of activated T8 cells. These experiments suggest the existence of stage-of-disease related suppressor cells in leprosy which appear to block the responsiveness of TH capable of responding to either specific or cross-reactive mycobacterial antigens. The mode of action of these Ts appears to be the inhibition of production of IL-2 and other lymphokines. Successful immunotherapeutic vaccination appears to overcome this block in the majority of patients.