[Cytomegalovirus infection in patients after kidney transplantation]. / Cytomegalovirová infekce u nemocných po transplantaci ledviny.

BACKGROUND: Cytomegalovirus disease (CMV) (active infection with various clinical and laboratory symptoms) in patients after renal transplantation (Tx) is an important diagnostic and therapeutic problem. Early CMV antigen pp65 is a structural protein of the antigenic type. It was discovered as a marker of activation of CMV infection. We decided to test the value of the method of early CMV antigen detection in the diagnosis of CMV disease in patients after renal transplantation (Tx). METHODS AND RESULTS: During a 10-month period 53 patients after Tx of the kidney were examined for the presence of early CMV antigen. Thirty-nine patients were followed up systematically, examined for the first time before Tx and then after regular one-week intervals. All patients were subjected to serological examination before Tx for the presence of antibodies against CMV, class IgG; in 41 during subsequent examinations also IgM antibodies were assessed. A method for the assessment of early CMV antigen was elaborated by cultivation of biological material in a tissue culture using monoclonal antibodies and indirect immunofluorescence. In the mentioned 53 patients a total of 363 examinations were made, mainly in blood and urine, 63 samples were positive Among the 39 patients followed up systematically early antigen was positive in 17 patients (44%). In three already before Tx. Positivity of the early antigen was first revealed on average 16 days (+/-4.2) after Tx and persisted on average for 13 days (+/-6.4). Positive early antigen was associated in the majority of patients with one or several clinical and laboratory symptoms, most frequently elevated body temperature. Comparison of early antigen and IgM anti-CMV were statistically evaluated by the chi sq. test, Fisher's test in contingency tables and by Student's non-parametric t-test. CONCLUSIONS: We assembled new experience with the method of detection of early CMV antigen in the diagnosis of activation of CMV infection. The correlation of the method with clinical and laboratory symptoms, incl. anti-CMV IGM antibodies, was statistically significant. By this method it is possible to detect CMV activation on average already on the 16th day after Tx which contributes in a significant way to the early diagnosis of the disease and facilitates the differential diagnosis of conditions during the early postoperative period.

[Hepatitis C virus antibodies in hemodialyzed patients]. / Protilátky proti viru hepatitidy C (HCV) u hemodialyzovaných nemocných.

Assessment of antibodies against the hepatitis C virus (HCV) in 629 patients treated by haemodialysis in 47 dialyzation centres revealed the probability of HCV infection in 196 of them, i.e. in 31%. In a group of 91 patients newly admitted for dialysis, antibodies against HCV were found in 24, i.e. in 26%. The percentage of anti-HCV antibody carriers in different dialyzation units varied between 6 nad 60.

[Cytomegalovirus antibodies: review of results of repeated examinations in 208 hemodialyzed patients]. / Protilátky proti cytomegaloviru: prehled výsledku z opakovaného vysetrení 208 hemodialyzovaných nemocných.

Within the framework of prevention of cytomegalovirus (CMV) transmission from infected kidney donors to non-infected recipients the author investigates as a matter of routine latent CMV infection by assessing the presence of serum IgG against pp150 CMV. Antibodies against this phosphoprotein of the viral matrix are found in more than 80% of haemodialyzed patients, i.e. potential recipients of kidneys (in our group 168 of 208 patients). During repeated examinations of 168 patients, positive as regards IgG against pp150 CMV, the author observed in 18 (11%) a temporary change to negativity. At the time of absence of serum IgG against pp150 CMV, however, in all patients except one serum IgG and/or IgG against some of the CMV antigen spectrum was found in the fibroblasts infected with cytomegalovirus. Specific IgM is an indicator of a change of the CMV infection from the latent to the active phase. The temporary loss of IgG against pp150 CMV may be an associated phenomenon.

[Epstein-Barr virus antibodies: review of results from repeated examinations in 208 hemodialyzed patients]. / Protilátky proti viru Epsteina a Barrové: prehled výsledku z opakovaného vysetrení 208 hemodialyzovaných nemocných.

Latent infection with the Epstein-Bar virus (EBV) is diagnosed as a matter of routine in haemodialyzed patients on the waiting list for renal transplantation by assessment of serum immunoglobulin G (IgG) against the nuclear antigen-1 (EBNA-1). Antibodies against EBNA-1 were detected in 163 of 208 patients (78.27%). Of these 158 entered the investigation already IgG positive against EBNA-1 and five converted later to positivity. Thirty-five of 50 patients who were IgG negative at the onset of the investigation, however, were IgG positive against the capsid antigen (VCA) EBV. Antibodies against EBNA-1 or against VCA were not detected during repeated examinations in the course of one an and a half years in 15 of 208 patients (7.21%).

[Elevated levels of cytomegalovirus antibodies and increased lymphocyte reactivity in patients with chronic renal failure]. / Zvýsené hladiny protilátek proti cytomegaloviru a zvýsená reaktivita lymfocytu u nemocných s chronickým selháním ledvin.

UNLABELLED: BASELINE: Cytomegaloviral infection is frequently encountered in patients with a reduced immunological reactivity, e.g. during chronic renal insufficiency; these patients have a markedly reduced cellular immunity and various lymphocytic anomalies. One of them is enhanced 3H-thymidine incorporation in non-stimulated lymphocyte cultures. The relationship between lymphocyte reactivity in vitro and activation of cytomegaloviral infection is the subject of the submitted paper. METHODS AND RESULTS: In 30 patients with chronic renal insufficiency the 3H-thymidine incorporation into lymphocytes and antibodies IgG and IgM against cytomegalovirus and the Epstein-Barr virus were investigated. In 15 patients the 3H-thymidine incorporation into lymphocytes in non-stimulated cultures was elevated, in 15 patients the incorporation was normal. All patients with an elevated 3H-thymidine incorporation the IgG serum levels against cytomegalovirus were markedly higher than in patients with normal incorporation. Serum IgM against cytomegalovirus was present only in patients with an elevated lymphocytic activity. Serum levels of antibodies against the Epstein-Barr virus (IgG) were identical in both groups. None of the patients had IgM against the Epstein-Barr virus. CONCLUSIONS: Enhanced 3H-thymidine incorporation into lymphocytes in uraemic patients reflects the defect of cellular immunity caused probably by elimination of some immunocompetent lymphocytes.

[Increased lymphocyte reactivity in patients with chronic kidney failure]. / Zvýsená reaktivita lymfocytu u nemocných s chronickým selháním ledvin.

Lymphocytes isolated from peripheral blood of patients with irreversible renal failure and cultivated in non-stimulated cell cultures incorporated significantly more 3H-thymidine and released more beta-2-microglobulin than lymphocytes of the patients' healthy relatives (possible kidney donors) and than lymphocytes of non-related blood donors.

Evidence for immunomodulatory properties of prolactin in selected in vitro and in vivo situations.

The relationship between neuroendocrine regulation and the immune system has recently become the subject of intense investigations. The pituitary secretes both immunostimulatory (growth hormone and prolactin) and immunosuppressive (ACTH) hormones, and is thus involved in the control of immune functions. The present work was aimed at the study of the immunoregulatory properties of prolactin in selected in vitro and in vivo model situations. Prolactin was found to enhance recovery of the receptor for sheep red blood cells (in vitro). Compared with control cells, incubation with prolactin and/or prolactin containing sera significantly enhanced the capacity of trypsin treated lymphocytes from the peripheral blood of healthy volunteers to form E-rosettes. Chlorpromazine stimulated prolactin release in males, and lactation stimulated prolactin release in females raised the number of large granular lymphocytes in peripheral circulation. Sera containing elevated prolactin levels stimulated the metabolic activity of peripheral neutrophilic leukocytes. These results suggest that prolactin may stimulate selective functions of cellular immunity, and that it is involved in interactions between the nervous, the hormonal and the immune systems.

Beta-2-microglobulin release in mixed lymphocyte cultures.

Significantly more beta 2-m is released in MLCs than in control cultures of cells from one donor or in cultures of pooled irradiated cells from several donors. Beta 2-m release increases with cultivation time and depends on the intensity of stimulation. It is highest in three-way MLCs and lowest in one-way MLCs when cells of haploidentical individuals are cocultivated.

Detection of antibodies to cytomegalovirus by immunofluorescence and enzyme immunoassay.

Immunofluorescence assay (IFA) and enzyme immunoassay (EIA) for detection of immunoglobulin M (IgM) to cytomegalovirus (CMV) correlated in 88.9%. All but one differences were due to negativity in IFA and positivity in EIA. The agreement between determination of immunoglobulin G (IgG) to CMV by IFA and determination of total antibodies to CMV by EIA was 89.9%. All the differences were due to a mild positivity in IFA and negativity in EIA. The advantage of EIA is in its objective colorimetric evaluation of test results excluding a subjective underestimation or overestimation of the reaction. In addition, EIA gives the result of IgM detection already in 6 hours, while IFA as late as in 24 hours.

[The effect of cytomegalovirus and Epstein-Barr virus infection on immunologic signs]. / Vliv infekce cytomegalovirem a virem Epsteina a Barrové na nekteré imunologické znaky.

Patients with active CMV or EBV infection characterized by the presence of specific serum IgM and/or high immunofluorescence in IgG, have higher levels of circulating immune complexes and a lower phagocytic activity of leucocytes than patients with latent CMV/EBV infections or not infected patients. In active infections at the same time the absolute number of lymphocytes with the surface sign CD8 increases. The amount of lymphocytes with differential antigens CD3 and CD4 does not change substantially. Also the levels of the third and fourth component of complement and C-reactive protein remain within the normal range.

[Decreased natural killer cell activity in health care personnel]. / Snízená aktivita prirozených zabíjecských bunek NK u pracovníku ve zdravotnictví.

Clinical immunology laboratory personnel have lower NK activity than those from administration and transport.

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.

Serum beta 2-microglobulin before and after renal and heart allotransplantation and in infection.

UNLABELLED: beta 2-microglobulin (beta 2-m) serum levels were significantly elevated in patients waiting for kidney transplantation. The levels decreased after successful transplantation, even when they were never as low as in a control group of healthy blood donors. A higher amount of serum (s-) beta 2-m was observed also in heart transplant recipients. A significant reincrease in s-beta 2-m occurred during rejection of renal allograft and during CMV and EBV infection. The levels of s-beta 2-m were higher in patients infected with CMV than in those infected with EBV. ABBREVIATIONS: beta 2-m = beta 2-microglobulin, s-beta 2-m = serum beta 2-microglobulin, CMV = cytomegalovirus, EBV = Epstein-Barr virus.

The immune profile of contact allergy patients.

Nonspecific parameters of antibody-related and cellular immune responses were studied in a group of 30 patients treated for manifest contact allergy to metals (chromium, cobalt, nickel) or non-metals (epoxy resin, rubber) confirmed in each of them by specific patch test positivity. In addition, every patient was tested for the pattern of skin reactivity to the Immunoskintest (USOL, Prague) antigens administered intradermally. The group of 20 metal allergy patients had decreased suppressor index values, enhanced formation of immediate active rosettes and an elevation of serum IgM and IgA immunoglobulins. The remaining 10 patients reacted by a reduced ability of neutrophils to exert phagocytic action on HEMA particles and a decrease in levels of alpha-2-macroglobulins in serum. This may help select a series of suitable immune parameters to be used as prediction tests in the secondary prevention of cases of occupational contact dermatitis.

Labelling of a T lymphocyte subpopulation with monoclonal antibody bound to colloidal gold. Correlation with the indirect immunofluorescence method.

A method of direct labelling of human lymphocytes CD8 with a monoclonal antibody bound to colloidal gold particles is described. Results of labelling of isolated lymphocytes and of cells of leucocyte concentrate from haemolysed blood correlated well with those obtained by indirect immunofluorescence. Owing to its simplicity and rapidity, the described method is particularly suitable for clinical diagnostics.

Natural killer cells and cytomegalovirus and Epstein-Barr virus infection in cardiac graft recipients.

The serum of 10 male patients undergoing heart transplantation was found to contain anamnestic antibodies (IgG) against cytomegalovirus (CMV) and Epstein-Barr virus (EBV). Patients with decreased natural killer (NK) cell activity displayed specific IgM against these viruses, a fact suggesting their activation and reproduction. Patients whose NK activity was within the range of that of healthy blood donors did not have serum IgM against CMV and EBV. Decreased NK activity and presence of specific IgM are associated with an increased number of LGL and monocytes, determined both by a monoclonal antibody (MEM-18) against their surface antigen, and morphologically.

CMV infection in patients with chronic renal failure and in those following transplantation.

Serum anti-CMV IgG was found in 62% of healthy blood donors. Patients in a chronic haemodialysis programme were seropositive in 80%, and kidney graft recipients in 93 to 100%, depending on the post-transplantation period. Median of the IgG titre was 1:10 in healthy persons, 1:80 in dialysed patients, and 1:40 in graft recipients. Anti-CMV IgG was never detected in healthy blood donors. IgM positivity was found in one of 30 dialysed patients, and in four of 27 kidney graft recipients. Compared with healthy blood donors, a higher percentage of CD 8 lymphocytes and LGL was found in dialysed patients exhibiting an anti-CMV IgG above 1:40. NK activity was impaired in all patients in the chronic haemodialysis programme.