Frequency and Aberrant CD Marker Profile of Acute Leukaemias.

OBJECTIVE: To determine the frequency of different types of acute leukaemia and their subtypes along with associated aberrant CD markers. STUDY DESIGN: Descriptive study. Place and Duration of the Study: Department of Immunology Armed Forces Institute of Pathology, National University of Medical Sciences, Rawalpindi, Pakistan, from November 2021 to October 2023. METHODOLOGY: All samples received for flow cytometric immunophenotyping with suspicion of acute leukaemia were included in the study. Cells were stained with fluorochrome labelled monoclonal antibodies against lineage-specific cluster of differentiation (CD) markers through a lyse-wash procedure. Acquisition and analysis were done using multi-parameter BD FACS Canto II Flow cytometer and BD FACS Diva software, respectively. Data were entered and analysed using SPSS v 23.0. RESULTS: Over a period of 2 years, a total of 1,115 suspected patients were tested for acute leukaemia. Among them, 728 (65.3%) were males and 387 (34.7%) were females, with mean age 28 ± 21 years, ranging from 1 week to 87 years. Among a total of 875/1115 (78.5%) diagnosed cases of acute leukaemia, AML was the most common leukaemia present in 408/875 (46.6%) patients followed by B-ALL and T-ALL in 384/875 (43.8%) and 70/87 (8%) patients, respectively (p = 0.5712). Aberrant CD markers were detected in 109/875 (12.5%) leukaemias (p = 0.0628). The most common aberrant CD markers in B-ALL were CD13 and CD33 present in 30/384 (7.8%) cases separately. Among AML and T-ALL most common aberrant CD markers were CD7 and CD33 present in 25/408 (6.13%) and 7/70 (10%) cases, respectively. CONCLUSION: Special consideration should be given to the presence of aberrant CD markers when assigning lineages to acute leukaemias. They may be important diagnostic, prognostic, and management tools for institution of immunotherapy. KEY WORDS: Aberrant CD markers, Acute leukaemia, CD Markers, Flow cytometry, Immunophenotyping.

HLA-DQ2 and HLA-DQ8 Alleles in Celiac Disease Patients and Healthy Controls.

OBJECTIVE: To compare HLA-DQ2 and HLA-DQ8 alleles between celiac disease patients and healthy control group. STUDY DESIGN: Observational cross-sectional study. PLACE AND DURATION OF STUDY: Department of Immunology, Armed Forces Institute of Pathology (AFIP), from April to December 2018. METHODOLOGY: Subjects were included: 100 celiac disease patients selected by non-probability consecutive sampling, and 100 healthy subjects. After collecting peripheral blood in EDTA tubes, chromosomal DNA was extracted and amplified, using sequence specific primers. Post-amplification electrophoresis was performed on two per cent agarose gel, followed by ethidium bromide staining; and specific band patterns were recorded under ultraviolet illumination to determine the HLA-DQ alleles. The subtypes of HLA-DQ2, i.e. HLA-DQ2.5 and HLA-DQ2.2 were also assessed. Frequency, percentage, mean and SD were calculated. Post-stratification Chi-square test was applied. RESULTS: The mean age of celiac disease group and healthy subjects was 14.79 ± 5.32 years and 14.71 ± 5.21 years, respectively. The frequency of HLA-DQ2 and HLA-DQ8 among celiac disease patients was 93% and 4%, respectively. Among HLA-DQ2 positive, HLA-DQ2.5 and HLA-DQ2.2 were found in 92% and 8%, respectively. Statistically significant difference (p <0.05) was observed between the celiac disease patients and healthy group. There was no significant difference observed among different age groups and gender (p >0.05). CONCLUSION: HLA-DQ2 detection reliably diagnoses celiac disease among all age groups and either gender. It can be used as an effective marker for early diagnosis of celiac disease instead of invasive procedures such as intestinal biopsy. The diagnosis can be pin-pointed by presence of HLA-DQ2.5. Key Words: Celiac disease, HLA-DQ2, HLA-DQ8, HLA-DQ2.5, HLA-DQ2.2.

Frequency of Panel Reactive Antibodies (PRA) among Renal Transplant Recipients and its Effect Modifiers.

OBJECTIVE: To determine frequency of panel reactive antibodies among renal transplant recipients and its effect modifiers. STUDY DESIGN:   A cross-sectional study. PLACE AND DURATION OF STUDY: Department of Immunology, Armed Forces Institute of Pathology from October 2016 to October 2017. METHODOLOGY: One hundred and sixty-two (162) patients, who were referred to Department of Immunology for pre-transplant workup for kidney transplantation of both genders and Pakistani nationality. Informed consents were taken and detailed history were recorded. Frequency and percentages were calculated for panel reactive antibodies, blood transfusion, pregnancy and previous transplant were noted and Chi-square test was applied. RESULTS: One hundred and sixty-two (162) patients including 141 males and 21 females were analysed and 48 patients (30%) were positive for panel reactive antibodies (PRA). Of 141 male patients analyzed, 35 were positive for PRA, which were about 25%. Twenty-one females were tested for PRA and 13 female patients were positive that is about 62% of the analysed population. Out of the total 141 males, 20 (14%) had blood transfusion and of these 11 (55%) were positive for PRA. Without history of transfusion, only 9 (7%) were positive for PRA. Out of 21 females, 10 were positive for blood transfusion, out of which 6 (60%) were positive for PRA. Without history of blood transfusion, 7 (64%) were positive for PRA. Out of 21 females, 20 had history of pregnancy. Out of whom, 13 (65%) were positive for PRA. Two patients (one male and one female) were with history of previous transplant and both were positive for PRA. CONCLUSION: A significant number of patients were sensitised with panel reactive antibodies waiting for renal transplant. The PRA was more common in recipients who were prone to effect modifiers such as pregnancy, blood transfusion and re-transplant. These risk factors were mostly present in combination, which also suggests their synergistic effects on PRA synthesis. Key Words: Blood transfusion, Effect modifiers, Panel reactive antibodies, Pregnancy, Kidney transplant, Re-transplant.