[Diagnostic value of determination of autoantibodies in gluten-sensitive enteropathy]. / Diagnostický prínos stanovení autoprotilátek u gluten-senzitivní enteropatie.

The study is focused on the immunopathological mechanisms of development of gluten-sensitive enteropathy (coeliac disease). It describes environmental factors and the role of autoantibodies and autoaggressive cells in the bowel inflammation. Attention is paid to the autoantibodies used in routine laboratory diagnosis of coeliac disease. The objective is a summary of rational diagnostic algorithms used in screening, diagnostics, treatment and dispensary care of patients with coeliac disease, especially with latent forms associated with other organ-specific immunopathological diseases. Exploration of anti-gliadin and anti-endomysial antibodies in diabetes mellitus type I were submitted. Furthermore, indications of these tests in the routine laboratory practice was analyzed.

[The chemiluminescence test in the laboratory evaluation of effects of immunostimulatory therapy]. / Vyuzití chemiluminiscencního testu pro laboratorní hodnocení úcinku imunostimulacní lécby.

BACKGROUND: Number of patients treated by general practitioners with various immunomodulatory drugs has recently increased. Effects of such medication on the immune system were not usually monitored. The aim of our work was to evaluate effect of selected immunomodulatory drugs on the phagocytic and metabolic activities of the phagocytes. METHODS AND RESULTS: 51 patients (18 males and 33 females) of the average age 36 years with repeating respiratory, mycotic and herpetic infections were investigated. Immunomodulatory treatment included: Decaris (Lavamizolum), Isoprinosine (Methisoprinolum), Imudon (Lysatum bacteriale mixtum), Biostim (Klebsiella pneumoniae), and Immodin (Leukocyti dialysati lyophylysatum). Before and after treatment all patients underwent basic immunological examination IgG, IgA, IgM, C3, C4 complement components, PEG, CD3, CD4, CD8 and CD19). Phagocytotic activity was estimated by means of FAGO MSHP test with HEMA particles and by chemiluminiscence test. Chemiluminiscence was measured using ML 3000 Microtiter Plate Luminometer (Dynex), 26 healthy individuals of the corresponding age were the controls. Results were statistically evaluated by Student's t-test. Significant increase of the cellular metabolic activity was found in Decaris and Immodin treated patients (P < 0.001). CONCLUSIONS: Chemiluminiscence test, which evaluates the metabolic activity of phagocytes, can be used for the accurate laboratory monitoring of the effects of some immunomodulatory drugs on the natural immunity of patients.

[Repeat kidney transplantation]. / Opakované transplantace ledviny.

BACKGROUND: The objective of the study was an analysis of results of repeated kidney transplantations (Tx2, Tx3) implemented during the first 29 years of activities of the Transplantation Centre of the Institute of the Clinical and Experimental Medicine in subjects with a different maintenance immunosuppression. METHODS AND RESULTS: The retrospective study pertains to 134 Tx2 and 17 Tx3 in 134 non-diabetic subjects: 43 of them had during Tx1 and Tx2 (1966-1981 and 1966-1985 resp.) immunosuppression on the basis of azathioprin (Aza, sub-group AA), 42 during Tx1 (1972-85), Aza, while during Tx2 (1984-85) immunosuppression on the basis of cyclosporin (CyA, subgroup AC) and 49 both during Tx1 and Tx2 (1985-93 and 1986-95 resp.) CyA (subgroup CC). Compared was survival of grafts by the actuarial method (with regard to all losses regardless of cause) by the end of the 4th year inside the subgroups (Tx2, vs. Tx1 and Tx3 vs. Tx2 in the same subjects) and between subgroups (Tx1 vs. Tx1 and Tx2 vs. Tx2 in different subjects). Moreover in paired investigations the survival of recipients and grafts after Tx2 was compared after immunosuppression on the basis of CyA with the same parameters after Tx1 in different subjects with the same immunosuppression, operated at approximately the same time (n = 81) and survival of subjects with Tx1 + Tx2 on the CC regime regardless whether the second grafts functioned at the time of the last examination, with survival of subjects after Tx1 where after graft failure Tx2 was not performed (n = 34). Prophylaxis with antilymphocyte globulins was not used. Survival of second and first grafts did not differ in any of the subgroups, third grafts survived at the end of the third year more frequently than second grafts (66 vs. 18%, p < 0.01). Second grafts in CC survived more than in AA (55 vs. 28%, p < 0.01). In the paired study Tx2 vs. Tx1 the survival of grafts and recipients was the same (88 vs. 89%, N.S. and 47 vs. 62% resp.), in the paired study Tx1 + Tx2 vs. Tx1 more subjects with Tx1 + Tx2 survived 10 years after Tx1 than subjects who did not have Tx2 (82 vs. 49%, p < 0.05). CONCLUSIONS: A further transplantation of the kidney after functional loss of the first graft is the method of choice: the mortality is low, the probability of several years' function is considerable and the prognosis as regards quality and length of life better than with regular dialysis treatment.

Kinetics of mixed lymphocyte reaction after blood transfusions.

The kinetics of mixed lymphocyte reaction (MLR) was studied in an unrelated pair of potential kidney graft donor and recipient. The recipient immunological reactivity to donor cells was specifically modulated by donor blood transfusions. Prior to transfusions, recipient MLR to donor lymphocytes expressed as per cent of the relative response (% RR) was 85.73. It dropped to 43.75 on the 21st posttransfusion day, and was as low as 19.16 on the 151st day (5th month). Next, a gradual increase in MLR occurred. The % RR increased to 46.15 in the 9th month and returned close to the pretransfusion level, to 78.30, in the 12th month. Transplantation of the kidney from the potential donor was not performed.

HLA molecule--cytoskeleton associations induced by cross-linking with specific antibodies.

Sera of patients with antibodies against HLA-typed panel, control human sera, anti-beta 2-microglobulin monoclonal antibody recognizing HLA class I molecules, anti-HLA class II monoclonal antibodies and control antibodies were used in indirect immunofluorescence study of living human peripheral blood lymphocytes and their cytoskeletons. Lymphocytes immobilized on concanavalin A-precoated glass slides enabling extraction procedures and repeated washings were found useful for such a study. If the lymphocytes were treated with human polyclonal anti-HLA antibodies only, or with two antibodies (i.e. with a specific monoclonal anti-HLA antibody and an anti-mouse Ig polyclonal antibody labelled with fluorescein isothiocyanate (FITC), the cross-linking of the surface molecules caused their resistance to the detergent effect. Murine monoclonal anti-HLA antibodies alone, bound to the cell surface molecules, formed immune complexes easily extracted with Triton solution. In such cases, the second anti-mouse Ig antibody labelled with FITC did not visualize specific molecules in the remaining matrix. No significant changes in the microtubules of cells treated with anti-HLA antibodies were observed.

Monitoring of immunological parameters before and after heart transplantation.

At the immunological examinations carried out prior to heart transplantation, it is crucial to check the compatibility in the ABO blood group system (regardless of the Rh factor) and to perform the cross match (which must be negative). In time pressure, cross match can be omitted provided the patient does not possess cytotoxic antibodies against a representative panel of lymphocytes. Other immunological tests before transplantation are of an auxiliary nature and should provide information whether the patient is a low or high responder, whether he does not suffer from an autoimmune disease or latent viral infection. In 37 out of 44 potential cardiac graft recipients, a strongly suppressed cell-mediated immune response was observed already in the pre-transplantation period. In the post-transplantation period, patients treated with corticoids and Cyclosporin A display a marked prolonged decrease in the absolute values of T4 positive lymphocytes. The absolute values of T3 and T4 lymphocytes represent a reliable parameter to assess the degree of achieved immunosuppression. Long-term monitoring did not reveal a significant association between the values of the T4/T8 index and the rejection episodes.

HLA antigens in systemic lupus erythematosus.

Forty-five patients suffering from systemic lupus erythematosus were studied in respect of their serologically defined HLA antigens. HLA-B8 antigen was found in 37-8% of patients as compared to 22% of controls. Individuals carrying the HLA-B8 antigen have a 2-15 times greater risk of developing systemic lupus erythematosus than those not carrying this antigen.

HL-A antigens in juvenile rheumatoid arthritis.

HL-A antigens were examined in sixty-two patients, thirty-one boys and thirty-one girls, diagnosed as having JRA. HL-A 27 was the only antigen with a significantly increased frequency. This increase concerned predominantly male patients in whom the disease developed at the beginning of puberty. In this group of sixteen boys, the polyarticular form was more frequent and tended to be associated with sacroliitis, while the rheumatoid factor was negative in all but one. The frequency of HL-A 27 in this group was 65.5%. The patients were also examined by the lymphotoxic test with anti H-2 alloimmune sera known to exhibit a high degree of association with some HL-A antigens. On JRA lymphocytes these associations were confirmed for HL-A 2 and anti-H-2f and for HL-A 27 and anti-H-2p. The strongest reactions were observed with lymphocytes from male patients around the age of puberty. These data indicate that steroid hormones influence the expressivity of some HL-A associated cell plasma membrane structures.