Doctoral Degrees (Paediatrics and Child Health)
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Browsing Doctoral Degrees (Paediatrics and Child Health) by Author "Coovadia, Hoosen Mahomed."
Item The antibody response to different measles vaccine strains given by the aerosol and subcutaneous routes to schoolchildren.(2003) Dilraj, Athmanundh.; Coovadia, Hoosen Mahomed.; Cutts, Felicity T.Abstract available in PDF.Item Hepatitis B virus-associated membranous nephropathy.(2002) Bhimma, Rajendra.; Coovadia, Hoosen Mahomed.Glomerulonephritis as an extra hepatic manifestation of chronic HBV infection has now been well documented [1,2,3,4,5]. HBV-associated nephropathy has been described in areas of both high and low endemicity [6]. In Africa HBV-associated nephropathy has been reported from the southern, central and northern regions [7,8,9,10,11]. In the southern African continent the prevalence of HBV-associated nephropathy appears to be higher than the rest of the continent [12]. In KwaZulu/Natal, South Africa, the prevalence of hepatitis B surface antigenaemia (HbsAg) in urban, rural and institutionalised children was reported to be 6.3%, 18.5% and 35.4% and the HBV exposure rates, as shown by the presence of any marker of HBV infection, 19.5%, 65.1% and 70.1% respectively amongst black children [13]. Prior experience of nephrotic syndrome (NS) and its association with HBV in black children, already published in a series of reports, showed HBV-associated nephropathy to be the commonest form of nephrotic syndrome among black patients in KwaZulu/Natal; membranous nephropathy (MN) being the commonest histological type reported [7,14]. The only other large series of HBV-associated nephropathy in southern Africa was from Cape Town of a large cohort of children, mainly of mixed ancestory (coloured), with a small number of black children [8]. There have been no other large studies of this condition amongst black children in Africa. We therefore undertook a series of studies to delineate the spectrum of this disease in black children with regard to the following: clinical presentation, laboratory findings, natural history, biosocial background, genetics (using HLA Class I and II antigens) as well as the impact of treatment and prevention by immunisation. We commenced these studies by reviewing our 20-year experience of 636 children with NS in Durban, South Africa for the period 1976- - 1995. Three hundred and six (48.2%) were blacks, 307 (48.2%) Indians and 23 (3.6%) were a mixed group (coloured); 91 (14.3%) could not be categorised and were excluded from the analysis. In black children, membranous nephropathy accounted for 43% of all cases of NS; 86.2% of these 306 children were associated with hepatitis B virus antigens [15]. This contrasts with the 2% - 5% prevalence of idiopathic membranous nephropathy reported in western countries [16]. We then proceeded to document the clinical features of this disease in black children. One hundred and thirty-three children with NS positive for HBV carriage were studied. In 70 patients the histological type was membranous; 46 of these 70 patients were followed up for a mean of 3.4 years (range 1-11). Spontaneous elimination of both HBsAg and HBeAg occurred in 10 (21.7%) of the 46 patients; 16 (34.8%) cleared HBeAg alone. Co-existing liver disease occurred in 18 (25.7%); hypocomplementaemia (low C3 and C4) in 22 (47.8%) and 5 (10.9%) of these 46 children respectively. Sixty-five (92.9%) of the 70 patients had normal renal function; 1(1.4%) impaired renal function; 3 (4.3%) chronic renal insufficiency and 1(1.4%) end stage renal disease at last hospital visit. Twelve (17.1%) of the 70 patients were in remission; all having cleared HBeAg. HBVMN was clinically indistinguishable from 24 children with idiopathic MN although biochemical characteristics were different. There were 23 patients with histological lesions other than MN. Forty patients with clinical, biochemical and serological findings similar to those with HBVMN and the other histological types, were unbiopsied. This report delineates the natural history of HBV infection in black South African children with NS, the majority of whom have MN. Disease remission in HBVMN parallels elimination of HBV antigens, particularly HBeAg. Comparison of HBVMN with idiopathic MN revealed clinically indistinguishable characteristics but unexplained biochemical differences [14]. Little is understood of the biosocial context in which HBV-associated nephropathy (particularly MN) develops. In the next two studies we evaluated HBV status and proteinuria in family members and household contacts of index children with HBVMN to test the hypothesis that HBV carriage and asymptomatic proteinuria are closely linked and may be causally associated. In the first of these two studies, thirty-one black children with biopsy-proven HBVMN were the index cases. One hundred and fifty-two family members and 43 black household contacts were the subjects of this study. We assessed HBV carrier status by testing for HBV antigens and antibodies using enzyme-linked immunosorbent assays (ELISA) and for HBV DNA by using slot-blot hybridisation and nested polymerase chain reaction. Sequencing of the precore HBV region of HBV was done in a subset of both index cases and subjects. Proteinuria was assessed by measuring the urinary protein: creatinine ratio. Seventy-two (37%) of the 197 family members and household contacts were HBV carriers, and 53 (27%) had a protein: creatinine ratio greater than the physiological limit (protein: creatinine ratio <0.2). Abnormal proteinuria was defined by a protein: creatinine ratio 0.2. Continuous data was compared using analysis of variance. Categorical data were compared using Chi-square test or Fisher’s exact test where appropriate. A probability of <0.05 was considered significant. The frequency of abnormal proteinuria was not significantly different in those with [22 (30.5%) of 72] or without [33 (32%) of 104] HBV carriage. This lack of association remained when carriers were classified into those who were HBsAg positive only and those with active viral replication (HBsAg and/or HBeAg and/or HBV DNA; p = 0.01). Family members were more predisposed to HBV carriage than household contacts, but abnormal proteinuria was present with equal frequency (p = 0.48). Age had a significant impact on proteinuria, with children less than five years being more likely to have abnormal proteinuria (p = 0.008). The prevalence of abnormal proteinuria in family members and household contacts of the index cases was more than in community-based controls. The 10 index HBVMN cases and 14 family members and household contacts that were tested all had HBV of genotype A. The results suggest that the family members and household contacts of children with HBVMN are at very high risk of HBV carriage; they also have asymptomatic proteinuria at a significantly higher rate than community-based controls. The HBV carrier status was not associated with proteinuria. This lack of association was a finding supported by peak prevalences of proteinuria in those under five years but no corresponding peak of HBV carriage. Proteinuria may indicate glomerular basement membrane dysfunction. Environmental and social factors may underpin development of these two disorders, but are insufficient to account for the index cases of HBVMN. The emergence of children with HBVMN from such households additionally depends on unidentified and possibly genetic factors [17]. In the second study of the biosocial background in which the HBV carrier-state with MN develops, we used the same subjects. One hundred and twenty-three unrelated individuals from the communities of the index cases, negative for HBV, served as controls. In this study, proteinuria was assessed using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and protein: creatinine ratios. Patterns of proteinuria on SDS-PAGE were classified as glomerular, tubular or mixed; IgG and haptoglobulin were suggestive of MN. Seventy-two (36.9%) of the 195 family members and household contacts were HBV carriers; 21 (29.2%) of these carriers had evidence of proteinuria using SDS-PAGE. Twenty-eight (41.2%) of the sixty-eight members of the study group who were HBV negative and 26.8% of the controls also showed proteinuria on SDS-PAGE. This lack of association between HBV carriage and proteinuria remained when controlled for gender and family relationship. Also, HBV was not protective against the development of proteinuria. Age was associated directly with a glomerular pattern of proteinuria (p = 0.007). Those having a pattern of proteinuria suggestive of MN were more likely to have an abnormal protein: creatinine ratio (p = 0.001). Ten (59%) subjects with a membranous pattern of proteinuria and 19 (47.5%) with a non-membranous pattern of proteinuria had microscopic haematuria. Such a pattern of proteinuria was not significantly different between subjects and community based controls (8.7% vs. 6.5%, p = 0.5). Environmental exposures in these subjects may be responsible for the proteinuria, which probably reflects underlying glomerular basement membrane damage. Discordance between the HBV carrier-state and patterns of proteinuria in the study group suggest that interaction between specifically vulnerable individuals and HBV group suggest HBV and MN may not be causally related or that it reflects exceptional interaction between specifically vulnerable individuals and HBV [18]. From the above two studies we inferred that the pathogenetic mechanisms by which individuals with chronic HBV infection develop MN are probably dependent on interactions between viral, host and environmental factors; some evidence suggests a genetic predisposition. We therefore undertook another two studies to explore HLA associations in black children with HBVMN. In the first of these two studies, thirty black children, age range 2 to 16 years, with biospy-proven HBVMN, were the subjects of this study. HLA A, B and C antigens were determined using a two-stage lymphocytotoxic test. HLA DRB1* and DQB1* typing was done using sequence-specific primers. HLA class I and II antigen frequencies of the study subjects were compared to controls that were randomly chosen healthy blood donors from the same population. HLA DQB1*0603 was increased in patients with HBVMN compared to controls (chi-square 13.65, RR 4.3). DRB1*07 and DQB1*02 were increased in frequency in the study subjects but failed to reach statistical significance. There was no significant difference in the frequencies of class I antigens in the study group compared to controls. This study is the first report of HLA associations in black patients with HBVMN in whom Class I and II antigens were determined using molecular methodology. It shows a high frequency of DQB1*0603 in black children with HBVMN compared to controls suggesting a possible genetic predisposition to the development of HBVMN [19]. Following our findings of an HLA Class II association in black children with HBVMN, we proceeded to determine if HLA DQB1*0603 predisposes to HBV carriage and development of abnormal proteinuria in the second study. We studied 70 family members of 14 children with HBVMN positive for HLA DQB1*0603 selected from the first study. Associations of HLA DQB1*0603 to HBV carriage and abnormal proteinuria were determined using the mean probability ratio (LOD scores). Forty-seven of the 70 (67%) family members were positive for HBV infection. Nineteen (27%) had abnormal range proteinuria. LOD scores in the study subjects with DQB1*0603 who were HBV negative vs. those with DQB1*0603 who were HBV positive was not significant (anti-log sum = 2.0559 and average 0.23). When a similar calculation was done for abnormal proteinuria, there were no significant findings (anti-log sum = 3.8587 and average 0.43). This lack of association between HLA DQB1*0603 with either HBV carriage or abnormal proteinuria in family members suggests that additional factors may play a role in predisposing children to chronic HBV carriage and the development of MN. We therefore conclude that the main effect of HLA DQB1*0603 which distinguishes HBVMN from family members is the degree of proteinuria which is a reflection of the severity of glomerular basement membrane damage in the latter [20]. In the next study we proceed to investigate the efficacy of Interferon alpha 2b (INTRON A ®) in the treatment of HBV-associated nephropathy in black children. Twenty-four black children with biopsy-proven HBV-associated nephropathy were recruited into the study during the period April 1997 to June 1999. Five defaulted treatment and were excluded from the primary analysis. IFN 2b was administered for 16 weeks. Response to treatment was defined as loss of HBeAg, decrease in proteinuria, and prevention of deterioration in renal and liver function. A control group of 20 patients was followed up for the same period. Ten (52.6%) of the treated children responded with clearance of HBeAg by 40 weeks. None cleared HBsAg. All responders showed remission of proteinuria, 90% maintained normal renal function and 1 (10%) showed improvement of renal function. HBV DNA levels decreased in this group. Nine patients did not clear HBeAg; none showed remission of proteinuria, 2 showed deterioration of renal function. Liver enzymes rose during treatment but subsequently declined irrespective of response to therapy. No serious side effects were encountered. Only 5% of controls showed spontaneous clearance of HBeAg, and none had remission of proteinuria. Black children with HBV-associated nephropathy show accelerated clearance of HBeAg with remission of proteinuria following treatment with IFN 2b. IFN 2b was well-tolerated [21]. We then went on to investigate the impact of HBV vaccination in South Africa over 6 years on HBV-associated MN. HBV vaccine has resulted in a decline in the incidence of HBV carriage and hepatocellular carcinoma in South East Asia. Vaccine efficacy in Africa has not been adequately assessed. King Edward VIII Hospital, Durban, South Africa, is the only tertiary referral centre for the province of KwaZulu/Natal for children with renal diseases. HBV vaccine was introduced into the Extended Programme on Immunisation (EPI) in April 1995; vaccine coverage rates between 1995-2001 for children for the first, second and third doses were 85.4%, 78.2% and 62.0% respectively. HBV status was determined using radioimmunoassay (1984 – 1991) or ELISA. MN was confirmed on renal biopsy. The hospital average annual incidence of HBVMN was compared pre and post-vaccination, and according to age groups. Between 1984 and 2001 there were 119 children with HBVMN; the mean age was 7 years (range 1 to 14 years) and 101(85%) were males. The average annual rate ratio (aRR) per 105 child population was 0.25. The aRR of 0.03 for the years 2000-2001, was significantly lower than the aRR of 0.22 during the pre-immunisation period (1984 – 1994) [p = 0.003; RR = 0.12 (95% CI: 0.03 – 0.5)]. The aRR in 2000-2001 for children 0 – 4 years (0.00) and 5 – 10 years (0.09) were significantly lower than in the pre-vaccination years (0.16 and 0.46, p = 0.01 and 0.02 respectively). Thus, HBV vaccine, even at low coverage for the full EPI schedule, reduced the hospital incidence of HBVMN by six years [22]. From this series of studies we concluded that prior to the introduction of the HBV vaccine into the Expanded Programme on Immunisation in Children, HBV-associated nephropathy, particularly MN was the commonest form of NS in black children. Several studies have suggested on the basis of epidemiological, clinical and immunological evidence a causal association between chronic HBV carriage and the development of nephropathy. In our present series of studies we have findings that lend further support to the causal association between HBV carriage and development of nephropathy, particularly MN, in black children. We have shown that genetic and other environmental factors may also play a role in determining the degree of proteinuria. Those children with abnormal range proteinuria less than the nephrotic range show no association with HBV carriage or genetic factors with regard to HLA linkage. The efficacy of interferon treatment in elimination of the HBV and abrogation of proteinuria following clearance of the virus (particularly the HBeAg) as well as the impact of routine HBV immunisation in preventing HBV carriage and subsequent development of nephropathy lends further support to our findings. The impact of viral load has yet to be investigated.Item Host allergic response variation in children with measles infection.(1977) Coovadia, Hoosen Mahomed.; Smythe, P. M.In many infections some patients recover while others die or are permanently disabled. These extremes in clinical outcome may be determined as much by the capacity of the host to eliminate the infecting agent as by the antigenic load on the individual. Children with measles who do not recover may succumb to acute complications (mainly respiratory) or chronic disease (respiratory and neurological) may develop. Analysis of immunological function antedating any of these final events would assist in understanding their pathogenesis and possibly aid in management. In order to achieve t h i s , immunological responsiveness was at f i r st studied in 24 children with acute measles and compared with that in 20 children with established chronic post measles chest disease investigated 6 - 1 6 weeks after appearance of the rash. The immunosuppressive effects of acute measles were extensive. Total white cells were reduced and this reduction was accounted for entirely by lymphopenia which was equally expressed among the major lymphocyte sub-populations studied; the function of T cells, assessed by radioisotope incorporation into phytohaemaggiutinin-transformed lymphocytes and by delayed skin hypersensitivity to dinitrochlorobenzene, was depressed. Serum IgA was reduced in acute measles patients. In contrast there was a relative sparing of the measured indices of immunity in patients with chronic post measles chest disease, with the major defect being an impaired delayed hypersensitivity reaction to dinitrochlorobenzene. There were minor alterations in complement components in both groups of patients with the evidence suggesting minimal utilisation of the alternative pathway in acute measles and classical pathway in chronic patients. High levels of heterophile antibodies to sheep red blood cells were detected in patients with chronic chest disease. (11) The results suggested that the conditions for chronicity of pulmonary disease in measles were unlikely to be determined by persistent abnormalities in the immunopathological factors enumerated, most of which were normal in chronic patients. It was not possible to interpret the findings of defective delayed hypersensitivity and complement components in patients with chronic chest disease as being either the cause or the effect of chronicity. The latter findings would have to be compared with results in children who had recovered from measles studied six weeks after onset of rash. An attempt was made to resolve this problem. Twenty-two children with measles were studied in the acute stage of the rash and six weeks later and results compared with matched controls. The above findings in acute measles were confirmed: the total lymphocyte count and major lymphocyte sub-populations were significantly below control values. At six weeks the B cell and Null cell counts were s t i ll significantly diminished. The function of T cells assessed by radioisotope uptake by phytohaemagglutinin-stimulated lymphocytes and by delayed skin hypersensitivity reaction to dinitrochlorobenzene was impaired during the acute stage and this persisted for six weeks. No important abnormalities were detected in serum immunoglobulins and complement components. Partial reversal of immunological suppression caused by measles was therefore demonstrated at 6 weeks after the appearance of rash. Demonstration of a persistently defective delayed hypersensitivity in those who recovered made i t unlikely that this anergy was important in the development of chronicity. Complement abnormalities were similarly unrelated to progression to chronic lung damage. ( Children who recovered, when studied at six weeks, appeared to be worse o f f immunologically than those with established chronic chest disease following measles. Children with chronic chest disease were studied 6 - 1 6 weeks after onset of rash, by which time the partial reversal of immune deficiency, noted at 6 weeks, would be complete. Among the group of children studied during the acute rash of measles there were five who subsequently died and one who progressed to chronic chest disease. Results in these six children were compared with those in six age-matched children who recovered from measles within a week. In the children who subsequently died or developed chronic pneumonia, immunosuppression was more pronounced during the acute rash ( i . e ., 3 - 2 0 days before death) than in the children who recovered. The absolute total lymphocyte count (T and B cells) was s i g n i f i c a n t ly lower in those who died or developed chronicity. Mean serum C, was also lower in this group. There were no significant differences between the two groups for total white c e l l s , neutrophils, Null c e l l s, cells with both T and B cell markers, other complement factors, serum immunoglobulins and phytohaemagglutinin stimulation of lymphocytes. The total lymphocyte count in a further nineteen patients with measles who had died, studied retrospectively, was s i g n i f i c a n t l y lower than that in twenty-seven patients with measles who recovered. Children whose outcome was poor generally had absolute lymphocyte 3 counts below 2000 cells/mm whereas those who recovered had values above this level. (iv) Therefore children who w i l l die or develop chronic chest disease can be often distinguished, within two days of the appearance of the rash, from those who w i l l recover. In order to test the v a l i d i t y of this conclusion based on results obtained from a small sample the study was extended so as to increase the number of patients with measles who had severe lymphopenia (< 2000 cells/mm3). Seventy seven per cent of 30 children who had severe lymphopenia within 2 days of appearance of rash f a i l e d to recover: 30% died from pulmonary complications within a few days to two months of the onset of the exanthem while 47% developed chronic lung damage. This was s i g n i f i c a n t ly worse than the outcome in 30 children with lymphocyte counts above 2000 3 cells/mm , of whom 67% recovered, 33% developed chronic chest disease and none died. Persistence of severe lymphopenia (which was due to reduction 3 in both T and B cells) in those with i n i t i a l counts below 2000 cells/mm , for at least fifteen days after onset of rash, remained a good predictive index of morbidity and mortality. Reversal of immunoparesis in those with i n i t i a l severe lymphopenia was slower and less complete 42 days from the appearance of the rash in children who subsequently died or progressed to chronicity than in those who recovered. All patients who died f a i l ed to produce an adequate or sustained antibody response to measles. The results of these studies suggest that long term pulmonary and possibly neurological sequelae of measles are probably due to a transient widespread immunoparesis during early measles with persistent defects in specific immunity to measles and probably other viruses, whereas recovery is due to less severe effects of shorter duration. (v) In order to answer the question why some children do badly and others well after measles, studies on the HLA frequencies and measles antigen load have been undertaken in children with severe lymphopenia. Results of viral load are inconclusive and those of HLA suggest a trend towards histocompatibility linked genetic susceptibility to the development of severe lymphopenia in measles associated with HLA AW32. The therapeutic implication of these studies is that children with measles who are at risk for death and chronic disease can be identified early in the disease and intervention at this stage may reverse the severe immunosuppression which leads to rapid demise or modify the immunopathological changes progressing relentlessly in some cases to permanent lung and brain damage and occasionally to death.Item Human immunodeficiency virus-1 infection and the acquired immunodeficiency syndrome in African children : natural history from birth to early childhood.(1999) Bobat, Raziya Ahmed.; Coovadia, Hoosen Mahomed.Background: in 1987, the first child with HIV-1 infection was identified in the paediatric wards at King Edward VIII Hospital in Durban. This made paediatricians aware that the epidemic had spread to the children of KwaZulu/Natal. Although information on transmission and natural history was becoming available from developed countries, little was known about the disease in developing countries. It was important to determine transmission rates and disease patterns in the local population, in order to appropriately counsel women, and for management of infected infants. In addition, with resources for laboratory diagnoses being limited in developing countries, much emphasis had to be placed on clinical findings for identification of infected children. In 1989, a retrospective analysis was made of the HIV-infected children seen over a 2-year period, between 1987 and 1989. Nine such children were identified and their clinical and biochemical features were described. It was concluded that HIV infected children presented with an identifiable pattern of signs, fairly similar to that described for children in industrialised countries. With these findings, a prospective study was undertaken, to determine the vertical transmission rate, the factors affecting this rate, and natural history of vertically transmitted I-IIV-1 infection. ix KwaZulu/Natal, being at the epicentre of the epidemic in South Africa, was a natural site for the study. Patients and Methods: a trained research worker was placed in the antenatal clinic at King Edward VIII Hospital for the specific purpose of educating, counselling, and testing of all women attending the clinic. Women attending the clinic for the first time in the index pregnancy were offered HIV testing if informed consent was obtained. Blood for HIV serology was drawn at the same time as sampling for the obligatory syphilis serology. The acceptance rate for sampling was > 95%. The majority of the women attending the clinic were black, and first attendance was generally late, into the third trimester. The same research worker was responsible for post-test counselling which was offered to all the women, not only those who tested positive. This research worker was also responsible for obtaining maternal consent for entering the newborn infant into the study. All newborn infants were seen within 48 hours of birth. At this time they were examined, growth parameters were recorded, and initial blood samples taken. These infants were then followed-up at 1 month, 2 months, 3 months, then at 3-month intervals up to 18 months, then at 6-month intervals. At each visit, a thorough clinical examination was performed, growth measurements taken, and development assessed. Record was made of any interim illness and visits to health centres, and of hospital admissions. Method of feeding was note& and details on immunisation obtained from the child's immunisation card. The children received all the x routine childhood immunisations according to the national regimen, based on WHO recommendations. Mothers were asked to bring the child to the follow up clinic for any problem, so that episodes of illness would not be missed. The women were reimbursed for transport costs to encourage follow up visits. Calculation of transmission rate and classification of infection status were made according to the recommendations of the Ghent workshop. Children were regarded as infected if they were antibody positive at 18 months or had an HIV related death. They were classified as uninfectd if the antibody test was negative at 9 months of age. Those infants who were lost to follow up before the age of nine months whilst still antibody positive and those whose cause of death could not be determined, were classified as indeterminate. The diagnosis of AIDS was based on the WHO criteria. Blood samples were taken at birth, at age one and three months, then at three month intervals to 18 months; thereafter at six month intervals. Sera were tested for HIV1 antibodies by a commercial enzyme-linked immunosorbent assay,ELISA. Samples that tested positive were confirmed by two tests, a Roche Elisa and by an immunoflourescent assay (IFA). A sample was regarded as being positive if both the second ELISA as well as the IFA or the Western Blot tested positive. xi Results: between October 1990 and March 1993, 234 infants and their 229 mothers were entered into the study. Those who did not attend a single follow up after birth were excluded from the study. The final cohort comprised 181 infants, of whom 48 were classified as infected ( including 17 deaths); 93 not infected, and 40 as indeterminate ( including 8 deaths). Maternal Data: about 60% of the mothers were under 30 years of age and were multiparous; 18% tested positive for syphilis serology; 22.9% were anaemic during pregnancy, and 37% were delivered by caesarean section. Most women lived in urban areas, and 16% chose to bottle-feed exclusively. Vertical Transmission Rate and Factors affecting this Rate: the median vertical transmission rate was 34%, (95% confidence intervals, CI 26%-42%). This figure is similar to that found in most parts of Africa, but much higher than those for Europe and USA. The maternal factors found to be associated with an increased risk of transmission were vaginal deliveries and a low haemoglobin level during pregnancy. Breastfeeding, Transmission, and Outcome: breastfeeding was found to have an increased risk of transmission, by 15 % (CI 1.8-31.8). On assessing growth and morbidity, it was noted that breastfed infants were not protected against such common childhood infections as pneumonia and diarrhoea, and that failure to thrive occurred with equal frequency in both those breastfed as well as those receiving artificial feeds. Newborn Data: when comparing newborn data between those infants who were subsequently found to be infected with those who were uninfected, it was found that there were no major differences between these groups with regard to growth parameters and neonatal complications. However, those infants with rapidly progressive disease (those who died within 24 months), were noted to have lower mean birth weights and lengths, a higher frequency of low birth weights, and tended to have more neonatal problems. Clinical Manifestations: the first differences between the infected and the uninfected infants generally manifested from about 3 months of age. HIV infected children were identifiable by higher frequencies of thrush, lymphadenopathy, skin rash, and hepatosplenomegaly in the early stages, and later on with a higher tendency to neurological and developmental abnormalities, as well as of diarrhoea. Pneumonia was found with equal frequencies in both the infected and uninfected children. The HIV infected child could be distinguished fairly early in life by the combination of the manifestations described above. Progression to AIDS: AIDS was diagnosed in 44% of all the infected children during the study period. Ninety five percent of these children were identified by 12 months of life, showing a rapid progression of the disease Longitudinal Growth: when longitudinal growth parameters were analysed in this cohort, it was found that HIV infected children were stunted from as early as 3 months of age, and remained below the international standards into early childhood. Infected children were also found to be malnourished (i.e. weight for age below international means), from an early age, and this persisted throughout early childhood. Of note, the uninfected childrens' weights, although comparable to international means initially, dropped after the first year of life. However, both groups did not have significant wasting, when compared to international means. Mortality: there were 25 known deaths during the study period. Of these, 17 were classified as HIV-related, and 8 as indeterminate. The mean age at death was 10.1 months, with 83% of all the HIV-related deaths occurring within the first year of life. The commonest diagnoses at the ti me of death were diarrhoea, pneumonia, and failure to thrive; also, thrush was common, as were neurological abnormalities.Item Neutralizing antibody responses and viral evolution in a longitudinal cohort of HIV subtype C infected antiretroviral-naïve individuals.(2011) Archary, Derseree.; Coovadia, Hoosen Mahomed.; Ndung'u, Peter Thumbi.Background: HIV-1 envelope (Env) diversity is arguably the most significant challenge for the development of an efficacious vaccine. An ideal vaccine would elicit the production of broadly neutralizing antibodies (nAb), capable of retaining potent activity against a diverse panel of viral isolates. The evolutionary forces that shape the diversity of envelope and ensuing nAb responses are incompletely understood in HIV-1 subtype C infection, the dominant subtype globally. Therefore there is an urgent need to define the patterns of envelope diversity, determine the correlates of immune protection and to discover subtype C immunogens in order to develop a globally relevant vaccine. Methods: We applied the single genome sequencing strategy to study plasma derived viruses from four slow progressors and four progressors over a median of 21 months between study entry and study exit. The participants‘ samples were from the Sinikithemba cohort of antiretroviral therapy-naïve chronically infected individuals and were termed slow progressors or progressors based on CD4 T-cell counts and viral loads over two years. We analyzed env sequence diversity, divergence patterns and envelope characteristics across the entire HIV-1 subtype C gp160. We studied the evolution of autologous nAb (AnAb) and heterologous nAb responses in order to test the hypothesis that slow disease progression is associated with more potent autologous or heterologous nAb responses. Furthermore, genotypic env characteristics were correlated to potency of neutralization in order to understand possible differences in nAb responses with divergent rates of disease progression and to describe genotypic differences associated with differential nAb potencies. In addition, the binding affinities of HIV-specific immunoglobulins (IgGs) and the affinities of the IgGs to various Fcγ receptors (both activating- FcγRI, FcγRIIa, FcγRIIIa; inhibitory- FcγRIIb) were assessed. These binding affinities were used as a surrogate for the recruitment of effector functions of cells of the innate immune system e.g. macrophages or natural killer cells to initiate antibody-dependent cell-mediated cytotoxicity (ADCC) or antibody dependent cell-mediated viral inhibition (ADCVI) and these were correlated to markers of disease progression namely CD4 T-cell counts and viral loads. Results: Intra-patient diversity was higher in slow progressors for regions C2 (p=0.0006), V3 (p=0.01) and C3 (p=0.005) compared to progressors. Consistent with this finding, slow progressors also had significantly increased amino acid length in V1-V4 with fewer potential N-linked glycosylation sites (PNGs) compared to progressors (p=0.009 and p=0.02 respectively). Similarly, in progressors, the gp41 region was significantly longer and had significantly fewer PNGs compared to slow progressors (p=0.02 for both parameters). Positive selection was prominent in regions V1, C3, V4, C4 and gp41 in slow progressors, whereas in progressors, it was prominent in gp41. Signature consensus sequence differences between the groups occurred mainly in gp41. Neutralizing antibodies (nAb) evolved over time in progressors, as evidenced by significantly higher nAb IC50 titers to baseline (study entry) viruses when tested against study exit time-point plasma compared to contemporaneous responses (p=0.003). In contrast, slow progressors‘ nAb titers did not differ significantly between study entry and study exit time points. nAb IC50 titers significantly correlated with amino acid lengths for C3-V5 (p=0.03) and V1-V5 (p=0.04) for slow progressors and V1-V2 for progressors (p=0.04). Slow progressors and progressors displayed preferential heterologous activity against the subtype C panel. There were no significant differences in breadth of responses between the groups for either subtype A or C. Neutralization breadth and titers to subtype B reference strains however, was significantly higher in progressors compared to slow progressors (both p<0.03) with increasing nAb breadth from study entry to study exit in progressors. Progressors had cross-reactive neutralizing antibodies that targeted V2 and V3. Binding affinities of non-neutralizing antibodies to HIV-specific gp120, gp41 and p24 and to activating and inhibitory Fcγ receptors (FcγRs) were similar in both groups. However, in slow progressors, CD4 T-cell counts correlated inversely with antibody binding affinity for the activating FcγRIIa (p=0.005). Conclusions: These data suggest that separate regions of Env are under differential selective forces, and the heterogeneity of env diversity and evolution differ with HIV-1 disease course. Single genome sequence analysis of circulating viruses in slow progressors and progressors indicate that diversity, length polymorphisms, sites under positive selection pressure, and PNGs consistently map to specific regions in Env. Cross-reactive neutralizing antibodies targeting epitopes in V2 and V3 indicate that nAb breadth may be dictated by a limited number of target Env epitopes. Certain key N-linked glycosylation sites were shown to be crucial for antibody neutralization. The potencies of autologous nAbs were directly affected by the amino acid lengths in certain regions of Env gp160 and by the numbers of PNGs. Target vaccine immunogens may have to be given over long periods of time and may have to include multiple subtype immunogens to elicit the production of potent, broad cross neutralizing antibodies with high binding affinity. Overall, the data suggest that neither nAbs nor non-neutralizing antibodies could be directly associated with disease attenuation in this cohort of chronically infected individuals. However, continuous evolution of nAbs was a potential marker of HIV-1 disease progression. Further studies on larger cohorts to identify people with potent nAbs and to identify specific targets of these antibodies are needed. Furthermore studies of non-neutralizing antibodies in HIV-1 infection using functional assays will be required in order to determine their role in HIV-1 pathogenesis.Item The role of immunoregulatory cells in healthy and sick African children.(1987) Kiepiela, Photini.; Coovadia, Hoosen Mahomed.Abstract available in PDF.Item Studies on the mechanisms of proteinuria in kidney diseases of childhood.(1994) Ramjee, Gita.; Adhikari, Miriam.; Coovadia, Hoosen Mahomed.Abstract available in PDF.