Detection of Anti-HLA Antibodies by Flow Cytometer

Lives of patients with solid organ failure depend physically, emotionally, and economically on others. Improvement in organ transplantation is one of the most important medical breakthroughs of the twenty-first century. Being healthy upon organ transplantation is the second chance to live the life. This is frequently observed in heart-, lung-, and liver-transplanted patients. For instance, upon kidney transplantation, dialysis dependence terminates and life quality of the patients increases. The major difficulty in organ transplantation is the low number of organ donation. Thus, the number of patients in the waiting list for the cadaveric transplantation increases day by day. Under these limited circumstances, required conditions should be further provided for the long survival rates of recipients with allogeneic graft without any problem. Human leukocyte antigen (HLA) tissue typing and anti-HLA antibodies produced before and after the transplantation adversely affect the graft survival and thus the survival of an individual. Investigation of pretransplantation immune status of recipients is significant. Particularly, donor-specific anti-HLA antibodies determine early and long-term graft survival. Flow cytometer is one of the most important devices used in anti-HLA antibody detection and also for other clinical and scientific purposes. Compared to conventional methods, it supports transplantation clinics due to its high sensitivity and specificity. The use of flow cytometer dependent methods in transplantation field increases progressively

Flow cytometric evaluation of T and B lymphocyte percentage in chronic kidney disease

Introduction. T and B lymphocytes play crucial roles in adaptive immunity. These cells are negatively affected in multiple disorders, including chronic kidney disease. The purpose of this study was to compare T and B lymphocyte ratios between patients with chronic kidney disease and healthy controls. Methods. In this study, we evaluated the percentages of patient and donor (healthy control) lymphocytes referred to our laboratory between 2012 and 2014. In total 103 patient-donor couples were tested by the FCXM method. CD3-PerCP and CD19-PE monoclonal antibodies were used in order to differentiate T and B cells, respectively. T and B cell percentages of the participants were statistically compared. Results. The mean age of the investigated patients and donors was 36.3 ± 13.7 and 46.2 ± 12.4 years, respectively. Of the studied patients, 45.6% and 54.3% were female and male, whereas 54.3% and 45.6% of donors were female and male, respectively. In the investigated group, 42 patients were preemptive, 45 subjects were treated with haemodialysis, and 16 individuals were on peritoneal dialysis. T and B lymphocyte percentages in the healthy group were higher than in patients with chronic kidney disease. However, the difference reached statistical significance only for T lymphocytes (p < 0.05). The percentages of total lymphocytes, and T and B lymphocytes in patients treated with haemodialysis were numerically lower than in those on peritoneal dialysis. In addition, we found that patients with chronic kidney disease had lower concentrations of haemoglobin and albumin than healthy controls. Conclusion. This study suggests that patients with advanced chronic kidney disease have lower rates of lymphocytes that healthy controls. This fact may at least partially explain impaired immunity in this setting. However, our findings require confirmation and detailed investigation of underlying mechanisms in further studies.

Böbrek nakli olan hastalarda nakil sonrasında oluşan anti-HLA antikorlarının saptanması

Bu çalışmada İzmir Tepecik Eğitim ve Araştırma Hastanesi’nde kadavra ve canlı donörden böbrek nakli olan hastaların nakil sonrasında haftalara göre toplanan serum örneklerinde hastanın grefte karşı immünolojik cevabının anti-HLA antikor mevcudiyetine bakarak değerlendirilmesi amaçlanmıştır. Bunun için 31 hastanın böbrek nakillerinden sonra bir yıl boyunca 1. gün, 1. hafta, 2. hafta, 4. hafta, 12. hafta, 24. hafta ve 52. haftalarda toplanmış olan serumlarında anti-HLA antikor tarama ve tanımlama testleri flow sitometri yöntemiyle gerçekleştirilmiştir. Bu yöntemle değerlendirilemeyen hastalar ise Luminex yöntemiyle doğrulanmıştır. Bu hastaların %12,9’u sadece sınıf I, %6,45’i ise sadece II PRA pozitif bulunmuştur. Hastaların hiçbirisi PRA sınıf I ve II pozitif bulunmamıştır. Hastaların %87,1’i PRA sınıf I negatif bulunurken, %93,5’i PRA sınıf II negatif bulunmuştur. Donör yaşı ile hastaların kreatin değerleri arasında anlamlı bir ilişki bulunmuştur (p<0,001). Hastaların yaş aralıklarına göre GFH istatistiksel açıdan değerlendirildiğinde aralarındaki ilişki anlamlı bulunmuştur (p<0,01). Bu çalışmadaki hastaların yaklaşık ortalama GFH’si 62,4±27,5 (ml/dk/1,73m2) ve ortalama kreatinin değeri 1,4±0,5 (mg/dL) olarak bulunmuştur. Ortalama GFH değerleri ile kreatinin değerleri arasında anlamlı bir ilişki bulunmuştur (p<0,001). Greft ve hastanın doku uygunlukları açısından değerlendirildiğinde canlı donörden nakil olan hastalar için en fazla görülen uyum 1A1B1DR şeklinde (%41,8, n=7) ve kadavradan nakil olan hastalar için en fazla görülen uyum 1B1DR (%28,57, n=4), 1B2DR (%28,57, n=4) ve 1A1B1DR (%28,57, n=4) şeklinde bulunmuştur. Greft naklini takiben 1. gün, 1., 2. ve 4. haftalarda greftteki uyumsuz antijenlere karşı antikor oluşmazken 12. haftadan sonra bazı hastalarda oluştuğu bulunmuştur