Is combined pretransplantation seropositivity of kidney transplant recipients for cytomegalovirus antigens (pp150 and pp28) a predictor for protection against infection?

OBJECTIVE: This study was aimed at detecting antibodies to the antigens which may contribute to protection against cytomegalovirus (CMV) infection after organ transplantation. MATERIALS AND METHODS: A total of 203 kidney transplant patients were enrolled in the study. Based on CMV antigenemia assay, 23 patients were antigen-positive and of the remaining 180 antigen-negative patients, 46 were selected as controls matched for age, gender and source of kidney. The 69 kidney recipients (KR) had CMV antibody due to previous infection and were followed up for a period of 6 months after transplantation for the development of active CMV infections by the antigenemia assay. Antibody responses to five CMV-related peptide antigens (pp65, gB, pp150, pp28 and pp38) were investigated by enzyme immunoassay and their presence was correlated with the results of the CMV antigenemia assay. RESULTS: Of the five CMV-related peptide antigens, only gB antigen showed response to the antibody in 10/23 (43.5%) antigen-positive patients and 9/46 antigen-negative patients and the difference was statistically significant (p = 0.048). On the other hand, there was no significant difference in antibody responses between the antigen-positive and antigen-negative KR to the other four CMV peptide antigens (p > 0.05). However, among the antigen-positive KR there was only 1 patient who had antibodies to both pp150 and pp28 antigen, while among the antigen-negative KR, 22 of 46 (47.8%) had the antibodies (p < 0.001). CONCLUSION: The findings suggest that the combined presence of antibodies against the pp150 and pp28 antigens may indicate a lower risk of CMV reactivation after kidney transplantation.

Oral valgancyclovir versus intravenous gancyclovir for cytomegalovirus prophylaxis in kidney transplant recipients.

BACKGROUND: Prophylaxis against cytomegalovirus (CMV) is a regular practice in organ transplantation. Oral valgancyclovir appears to be an interesting alternative to the usual intravenous form. PATIENTS AND METHODS: We prospectively compared the response of intravenous gancyclovir for 2 weeks (GAN; n=41) to oral valgancyclovir for 2 weeks (VAL2w; n=23) or 3 months (VAL3m; n=46) in kidney transplant recipients receiving induction immunosuppression. CMV antigenemia assay and/or polymerase chain reaction (PCR) were used for viral detection. Patients were followed for a minimum of 6 months posttransplantation. SPSS software was used for statistical analysis using a cutoff of significance as P<.05. RESULTS: There was no statistical difference in the demographic features among the study groups. However, human leukocyte antigen (HLA) match was better in the VAL3m group and the patients of this group received less ATG induction immunosuppression (41.3%) compared with the GAN group (100%). The incidence of acute rejection was not different among the study groups. There was a higher incidence of fever with positive CMV tests in the VAL2w group (P=.035) compared with the other groups, while leukopenia with a negative CMV test was significantly higher in the VAL3m group (P=.04). The incidence of CMV disease was higher in the VAL2w group (30.4%) compared with the GAN group (14.6%) or the VAL3m group (8.7%). Renal function was significantly worse in the VAL2w group at 3 and 6 months (P=.011 and .02, respectively). CONCLUSIONS: Three months oral valgancyclovir prophylaxis for CMV was a more effective regimen compared with intravenous gancyclovir for 2 weeks. Shorter courses were associated with a higher incidence of CMV infection and poorer graft function. Leukopenia observed in patients receiving valgancyclovir may be a drug-related side effect.

Herpesvirus antibodies and antigens in patients with cervical anaplasia and in controls.

Antibody activity to herpesvirus hominis type 2 (HVH-2) in 151 women cured of cervical carcinoma (60 in situ, 91 invasive) and in 106 controls differed significantly, especially between the in situ (73%) and control (17%) groups. Sera of 57 patients with cervical atypia showed an increased antibody activity to HVH-2 (58%), compared with that of the 57 matched controls (23%). Antibodies to cytomegalovirus (CMV) were detected more frequently in sera of women with atypia (61%) than in sera of women with cervical disorders other than atypia (42%) or in sera of healthy controls (33%). HVH antigens were present in cervical cells from patients with atypia and from matched controls. Not only exfoliated (imprints) but also cultured and cocultured cervical cells contained HVH antigens; there was no correlation between antigen positivity and antibody activity to HVH-2. Our data further supported the association between HVH-2 and cervical anaplasia and indicated that CMV may also be implicated in its etiology.

Herpes simplex virus-specific antigens in exfoliated cervical cells from women with and without cervical anaplasia.

By the indirect immunofluorescence technique the presence of herpes simplex virus-specific antigens was investigated in cervical cells of 530 women with normal cervical epithelium, 175 with bland disorders, 52 with dysplasias, and 38 with invasive cervical carcinomas of the uterine cervix. Antigens were present in 9% of samples from women with normal cervical epithelium; they were present in 41% of the samples from women with bland disorders, 61% of those from dysplasia patients, and 94% of those from invasive carcinoma patients. The testing of 3 consecutive imprints of 68 antigen-positive and 232 antigen-negative women at 6-month intervals revealed that, in cervical cells, herpesvirus-specific antigens persisted throughout the 1-year period of the follow-up.

BK virus nephropathy in renal transplant recipients in Kuwait: a preliminary report.

INTRODUCTION: BK virus nephropathy (BKVN) is a significant cause of graft loss among renal transplant recipients. The treatment outcomes of BKVN have been variably reported in the literature. PATIENTS AND METHODS: We prospectively investigated BKV infection and BKVN among a population of renal transplant recipients with suspected BKV infection. The 42 subjects who all had acute allograft dysfunction, were categorized in three groups: those with clinical, laboratory, and histological findings that did not suggest acute rejection, drug toxicity, or obstruction (group 1, n = 24); those with findings that suggested probable acute cellular rejection but did not respond to antirejection treatment (group 2, n = 10); and those whose renal histology suggested BKVN (group 3, n = 8). Polymerase chain reaction analysis was done to detect BKV DNA in urine and blood samples from each subject. BKV DNA was detected in 19 (45%) urine samples with 11 of these subjects (26.1% of total) having BK viremia as well. RESULTS: No evidence of BKVN was detected histologically in seven subjects with isolated BK viruria, while the others proved to be JC virus infections. Among the 11 subjects with BK viremia, eight had BKVN based on renal histology at the time of diagnosis with BKV infection, while the other three subsequently developed histological features of BKVN. BKVN developed after 5.3 +/- 2.5 (2 to 44) months after transplantation. The serum creatinine at time of BKVN diagnosis was 158.9 +/- 58 (87 to 285) micromol/L. All subjects were initially treated with a 50% reduction in immunosuppressive drug doses. Further decreases in immunosuppression were performed in all patients with close monitoring of renal function. All subjects were followed up for a of 18.2 +/- 5 (12 to 26) months. Two grafts were lost not due to BKVN, and one patient was lost to follow-up during this period. The latest serum creatinine in eight recipients is 113 + 20 (81 to 138) micromol/L, which is better than the renal function at diagnosis. CONCLUSION: The prevalence of BKVN in suspected BKV infection was 26%. Although the study period was short (30 months), BK viremia strongly correlated with BKVN, which seemed to be successfully treated with reduction in immunosuppression.

The suppressive effect of progesterone on lymphocyte cytotoxicity: unique progesterone sensitivity of pregnancy lymphocytes.

The effect of progesterone on the cytotoxic activity of lymphocytes obtained from healthy pregnant women, women with threatened pre-term delivery, healthy non-pregnant women and healthy male donors has been compared. The cytotoxic activity of lymphocytes from healthy pregnant women was significantly reduced by progesterone at concentrations present in the serum during pregnancy. In contrast, a 100-fold higher concentration of progesterone was required to diminish the cytotoxic activity of lymphocytes from women with threatened pre-term delivery and from healthy male donors. Individuals with lymphocytes of high and low progesterone sensitivity could be found amongst non-pregnant women. The results of investigations at the single cell level suggested that although progesterone did not inhibit the ability of the lymphocytes to bind to the target cells, it markedly reduced the target cell lysing capacity of the bound effector cells.

Progesterone as an immunologic blocking factor in human pregnancy serum.

The influence of estriol and progesterone on lymphocyte reactivity was examined by testing the cytotoxic effect on human embryonic fibroblast cells of non-pregnant women's lymphocytes incubated with different concentrations of estriol and progesterone and with complete and progesterone-depleted third trimester pregnancy sera. Progesterone, but not estriol, had a significant inhibitory effect on lymphocyte cytotoxicity at concentrations comparable to those present in pregnancy serum. 95% depletion of progesterone from pregnancy sera caused an 80% loss of inhibitory activity on lymphocyte cytotoxicity. These data suggest that the blood level of progesterone in pregnancy is sufficient to depress lymphocyte reactivity and that progesterone is responsible for the greater part of the serum inhibitory activity in at least the later stages of pregnancy.

Immunosuppressive effect of serum progesterone during pregnancy depends on the progesterone binding capacity of the lymphocytes.

Cytotoxic activity and progesterone binding capacity of the lymphocytes, together with serum progesterone concentrations, were determined in women with normal pregnancy or with a clinical diagnosis of threatened abortion or threatened premature labour. The lymphocytes of women with threatened abortion or threatened premature labour showed significantly higher cytotoxic activity (P less than 0.001) and significantly lower progesterone binding capacity (P less than 0.001) than did lymphocytes obtained from the healthy pregnant women. Significant inverse correlation was found between progesterone binding capacity and cytotoxic activity of the lymphocytes (P less than 0.001), but the progesterone concentration of the pregnancy serum appeared to have no influence on the other two parameters. The findings indicate that intact progesterone binding capacity of the lymphocytes is an essential factor for the manifestation of the blocking effect exerted by pregnancy serum on lymphocyte cytotoxicity in vitro.

Role of interleukin-12 in patients with dengue hemorrhagic fever.

Interleukin (IL)-12 has a broad range of activities including regulation of cytokine synthesis and selective promotion of Th1-type cell development. A shift from a Th1-type response to Th2-type has been suggested to be important in the pathogenesis of dengue hemorrhagic fever (DHF). This study was undertaken to investigate the possible role of IL-12 in this shift. A total of 76 patients with various grades of dengue illness and 21 normal healthy controls were tested for IL-12 levels in serum samples and IL-12 mRNA in their peripheral blood mononuclear cells. The results showed that the levels of IL-12 were the highest in patients with dengue fever (270+/-102 pg ml(-1)) followed by decreasing levels in the patients with DHF grade I (198+/-86 pg ml(-1); P<0.05) and DHF grade II (84+/-52 pg ml(-1); P<0.001). Neither IL-12 nor its mRNA could be detected in the patients with DHF grades III and IV. The cytokine appeared and reached peak levels during the first 4 days of illness, started to decline by day 5-8 and disappeared by day 9 onwards. The absence of IL-12 during severe illness and late phases of the disease may be responsible for the shift to a Th2-type response and thus for the pathogenesis of DHF.

Hantavirus-specific antibodies in rodents and humans living in Kuwait.

Hantaviruses are found in widely scattered areas of the world and are transmitted by inhalation of virus-contaminated aerosols of rodent excreta. The present study was undertaken in Kuwait to investigate the serological evidence for hantavirus infection in rodents and humans. Sera were collected from 283 wild rodents and 183 human subjects (46 Kuwaitis and 137 non-Kuwaitis). The rodent sera were investigated for the presence of antibodies against the Seoul and Puumala strains of the hantaviruses by enzyme-linked immunosorbent assay and immunofluorescence technique using the virus-infected Vero E6 cells. The findings showed the presence of anti-hantavirus antibodies in seven out of the 283 (2.8%) rodents. Antibodies against the Seoul strain were present in six (2.1%) and against the Puumala strain in three (1%) rodents. Further, it was observed that three out of 84 (3.6%) of the Rattus norvegicus and four out of 174 (2.3%) Mus musculus had anti-hantavirus antibodies. Two rodents belonging to species Mus musculus had antibodies against both strains of the hantaviruses. Out of 183 human sera, 13 (7%) were positive for hantavirus antibodies. Among the Kuwaitis 5/46 (11%) and among the non-Kuwaitis 8/137 (6%) were positive for the hantavirus antibodies. Antibodies to both Puumala and Hantaan strains were detected in Kuwaitis as well as in non-Kuwaitis. Although no human case of hantavirus illness has yet been reported in Kuwait, the serological evidence of infection suggests a constant vigil.

Correlation between CMV genotypes, multiple infections with herpesviruses (HHV-6, 7) and development of CMV disease in kidney recipients in Kuwait.

The possible correlation between cytomegalovirus, human herpesvirus types 6, 7 and cytomegalovirus-related clinical symptoms was studied in kidney transplant patients in Kuwait. Cytomegalovirus infection was diagnosed using the pp65 antigenemia assay. DNA of cytomegalovirus was detected by nested polymerase chain reaction (nested-PCR). PCR was also used to amplify the genes coding for structural proteins of human herpesvirus-6 (240 bp) and human herpesvirus-7 (186 bp). Glycoprotein B genotypes of cytomegalovirus were determined by restriction fragment length polymorphism. The average number of cells positive for cytomegalovirus pp65 antigen showed a steady increase with the severity of the cytomegalovirus-related symptoms. Furthermore, cytomegalovirus pp65 antigen positivity was significantly more frequent among recipients of cadaver kidney (45.5%) than among those who received live related kidneys (22.6%). Cytomegalovirus gB genotype 1 was detected more frequently (P<0.036) in recipients with live related donor kidney (38%) than in patients of cadaver kidney (13%). The genome of human herpesvirus-6 was detected at the same rate in patients with or without cytomegalovirus-related symptoms. However, the genome of human herpesvirus-7 was detected significantly more frequently (P<0.0001) in asymptomatic patients (41.7%) than in recipients with symptomatic cytomegalovirus infection (17%). We conclude that cytomegalovirus gB genotypes are not associated with the outcome of a cytomegalovirus infection in kidney transplant patients, that human herpesvirus-6 does not play a role in cytomegalovirus pathogenesis and that the role of human herpesvirus-7 in cytomegalovirus-related morbidity in kidney recipients remains unclear.

The prevalence of antibody to human parvovirus B19 in pregnant women in Kuwait.

Infection with parvovirus B19 during pregnancy is known to be associated with various fetal damage, such as aplastic anemia and hydrops fetalis. Our objective was to study the seroprevalence of parvovirus B19 in the obstetric population in Kuwait and to compare this with that in the adult population in other regions. Blood samples from 1047 pregnant women were used in this prospective study. Information regarding patient's age, parity, nationality and symptoms was obtained at the time of collection of the sample. Blood was tested for IgG and IgM antibodies specific for parvovirus B19 using the ELISA technique. The overall prevalence for IgG and IgM was 53.3 and 2.2%, respectively. IgG seropositivity was higher in non-Kuwaiti women, while IgM antibodies was more frequent in Kuwaiti women. A total of 17.4% of the acutely infected patients were symptomatic. Prevalence of parvovirus B19 infection in Kuwait is comparable with that in the other countries.

The mechanism of the inhibitory effect of progesterone on lymphocyte cytotoxicity: II. Relationship between cytotoxicity and the cyclooxygenase pathway of arachidonic acid metabolism.

In the previous paper we have demonstrated that progesterone-treated lymphocytes of healthy pregnant women produce a 34,000 MW protein that inhibits cytotoxic activity and prostaglandin F2 alpha synthesis. Since recently it has been shown that certain leukotrienes have a stimulatory effect on natural killer activity, in this study an attempt was made to determine whether there is a relationship between cytotoxicity and PGF2 alpha synthesis or if alterations in the values of these parameters are independent. Arachidonic acid increased cytotoxic activity of healthy pregnant women's peripheral blood mononuclear cells (PBMC) in a concentration-dependent manner. Exogenous arachidonic acid was able to counteract the blocking effect of the above-mentioned protein produced by progesterone-treated lymphocytes. To determine whether the products of the cyclooxygenase or the lipoxygenase pathway of arachidonic acid metabolism are responsible for increased cytotoxicity, both enzyme systems were blocked separately. Both indomethacin and the lipoxygenase inhibitor BW 755C reduced cytotoxicity in a dose-dependent fashion. However, the lipoxygenase inhibitor prevented prostaglandin synthesis to the same extent, or even more than indomethacin, in all concentrations used; so, its blocking effect cannot be considered as supportive evidence for the role of leukotrienes in cytotoxicity. On the other hand, lipopolysaccharide, with a selective stimulatory effect on prostaglandin synthesis, increased cytotoxicity. Lipopolysaccharide had no effect on progesterone-pretreated PBMC. The above data allow the assumption that besides leukotrienes, cyclooxygenase products may also increase cytotoxicity.

Increased NK activity is responsible for higher cytotoxicity to HEF cells by lymphocytes of women with threatened preterm delivery.

Recently we have shown that lymphocytes of pregnant women with threatened preterm delivery (risk group) exerted significantly higher cytotoxic activity to human embryonic fibroblast (HEF) cells than those of healthy pregnant women. The purpose of this study was to get information on the mechanism of this cytotoxicity. The possibility of prior sensitization to embryonic antigen was excluded, since no difference could be demonstrated between cytotoxic activity of lymphocytes obtained from women with two or more previous pregnancies and that of lymphocytes from never-pregnant women. For determining the effector cell type responsible for cytotoxicity, lymphocytes of 50 healthy pregnant women and those of 50 risk patients were tested in different cytotoxicity tests using HEF and K-562 target cells. The proportion of NK cells among lymphocytes was determined by counting large granular lymphocytes (LGL), IgG Fc receptor bearing cells, and cells positively stained by NK specific monoclonal antibody. Though no difference in the proportion of NK cells between the two groups was found, risk patients' lymphocytes were significantly more cytotoxic to K-562 target cells than those of healthy pregnant women. Investigations at the single-cell level made it obvious that this higher cytotoxic activity originated from increased target cell lysing ability of their lymphocytes, while their conjugating capacity did not differ significantly from that of lymphocytes obtained from healthy pregnant women.(ABSTRACT TRUNCATED AT 250 WORDS)

The mechanism of the inhibitory effect of progesterone on lymphocyte cytotoxicity: I. Progesterone-treated lymphocytes release a substance inhibiting cytotoxicity and prostaglandin synthesis.

Progesterone-treated lymphocytes (generator lymphocytes) of healthy pregnant women release a nondialyzable factor that inhibits both cytotoxic activity and prostaglandin F2 alpha synthesis of test lymphocytes. Production of this factor is blocked by protein synthesis inhibitors (cycloheximide and actinomycin D). Sodium dodecylsulfate polyacrylamide electrophoresis of the partially purified material revealed a main 34,000 MW protein band. Progesterone-treated lymphocytes of pregnant women showing clinical symptoms of threatened preterm delivery (risk group) failed to release this substance.

Cytomegalovirus prophylaxis with ganciclovir in kidney transplant recipients receiving induction antilymphocyte antibodies.

BACKGROUND: Cytomegalovirus (CMV) is one of the serious viral infections after organ transplantation, especially in patients receiving anti-lymphocyte antibodies. Prevention of the infection using antiviral chemotherapy (ganciclovir) has gained interest in the transplant community due to the availability of quantitative methods for viral detection and monitoring. METHODS: Forty-six CMV seropositive kidney transplant recipients were assigned to receive induction immunosuppression with anti-thymocyte globulin (ATG, Fresenius). Prophylactic intravenous ganciclovir was administered for 2 weeks at a dose of 5 mg/kg/d (adjusted to kidney function) starting from the day of surgery. Patients were monitored regularly for CMV infection or disease over 1 year posttransplant. The time to CMV manifestation, the number of antigenemia assay-positive cells, the clinical severity of infection, the incidence of acute rejection, the graft function, and the duration of hospital stay were evaluated. This group was compared to a historical matched control cohort (n = 37) transplanted earlier who did not receive prophylactic ganciclovir. RESULT: The incidence of CMV disease was significantly less among the prophylaxis than the control group (6/46 patients [13%] vs 16/37 patients [43.2%], P = <.004). The time to develop CMV manifestations was much longer in the prophylaxis group than in the control group (median 92 vs 32 days, P

A study of dengue imported to Kuwait during 1997-1999.

This study was carried out on sera from 210 patients in Kuwait in 1997-1999. All of the patients were suffering from febrile illness and had recently visited dengue- (DEN) endemic areas. The sera were screened for DEN virus by inoculation into cultures of the Aedes albopictus cell clone C6/36 (virus isolation) and by IgM capture ELISA (detection of DEN virus-specific IgM antibodies). In the cell cultures, DEN virus could not be isolated from any of the patients' sera. However, sera from 19 patients were positive for DEN virus-specific IgM antibodies. All these 19 sera were tested for the presence of DEN virus-specific RNA by reverse transcription-PCR (RT-PCR) using DEN virus types-common (consensus) primers. In addition, the type of DEN virus was identified by using type-specific primers in a semi-nested PCR. The results showed that two of the 19 patients were infected with DEN virus type 2. This report of 19 patients with serological evidence of DEN infection indicates that imported DEN is a real threat to Kuwait, a country non-endemic for DEN but with a large portion of the population vacationing in DEN-hyperendemic areas during the peak DEN season and then returning to Kuwait.

Seroprevalence of three emerging arboviral infections in Kuwaiti nationals.

Diseases caused by dengue, sandfly fever and hanta viruses pose a major health risk in many countries. We determined the threat of these arboviral infections through a serologic using enzyme linked immunosorbent assay (ELISA) based tests. Hantavirus-specific antibodies were also detected using immunofluorescence. Of 499 samples tested for dengue virus IgG antibodies l4% were as positive for dengue positive by all the ELISA tests. Among the 42 showing strong IgG reactivity, only 1 was positive for dengue virus IgM antibodies. All samples tested for IgG antibodies to sandfly fever virus were negative. Hantavirus antibodies were detected in 11% of the 46 samples from high-risk individuals. The low prevalences suggest that at present these infections are not a serious problem in Kuwait.

Cytomegalovirus genotypes gB1 and gH1 are the most predominant genotypes among renal transplant recipients in Kuwait.

BACKGROUND: The human cytomegalovirus (HCMV) is a common pathogen responsible for asymptomatic and persistent infections in healthy individuals. However, cytomegalovirus infections are a major cause of morbidity and mortality in immunocompromised patients, especially in recipients of solid-organ transplants and AIDS patients. METHODS: HCMV DNA from 42 patients who received kidney transplants between 2004 and 2008 were subjected to polymerase chain reaction and restriction fragment length polymorphism to identify HCMV gB and gH genotypes. RESULTS: HCMV gB1 and gH1 genotypes were the most the predominant HCMV genotypes (P < .05, P < .05, respectively). In addition, both HCMV gB1 and gH1 genotype were significantly more often associated with the development of fever with leukopenia and severe HCMV disease than other gB or gH2 genotypes. No significant differences were observed among viral loads between the HCMV genotypes among infected individuals. CONCLUSION: This study demonstrated the prevalence and role of HCMV genotypes in infection and disease in renal transplant patients in Kuwait.