Malawian Patients Research Articles

Inter‐ and intra‐person cytomegalovirus infection in Malawian families

Sequence polymorphisms in the gN and gO genes of cytomegalovirus (CMV) amplified from mouth rinse and urine samples of 19 Malawian patients with Kaposi's sarcoma (KS) and 58 of their first-degree relatives were investigated. CMV-DNA was amplified from 41 people (53%) from either the gN or gO region in at least one sample, from 14 people (18%) in both domains in at least one sample, and from 13 (17%) in either domain in both samples. Twenty-one (51%) were seropositive for human immunodeficiency virus-1 (HIV). Identical gN sequences were recovered from eight families and non-identical sequences in six, while identical gO sequences were found in three families and non-identical sequences in five. Five people, four of whom were children, each carried multitypic gN sequences or gO sequences. The findings are consistent with CMV spread along intra- and extra-household routes, and with multiple intra-host CMV infection.

Ocular disease in patients with TB and HIV presenting with fever in Malawi

To investigate ocular disease in Malawian patients with tuberculosis (TB) and HIV in presenting with fever, and to determine if indirect ophthalmoscopy is useful in the diagnosis of mycobacteraemia. A prospective study of all adult patients admitted with fever to Queen Elizabeth Central Hospital, Blantyre. All recruited patients had an ophthalmic examination, HIV tests, chest x-ray, sputum examinations, bacterial and mycobacterial blood cultures and malaria slide. 307 patients were recruited; 109 (36%) had TB, including 53 (17%) with mycobacteraemia; 255 (83%) had HIV and 191 (62%) had AIDS. Of the patients with TB 102 (94%) had HIV. Choroidal granulomas were found in four patients, all of whom had AIDS; three had disseminated TB with mycobacteraemia, and one had persistent fever but no other evidence of TB. Among the patients with AIDS, 32 (17%) had retinal microangiopathy manifest by cotton wool spots; one (0.5%) had signs of active cytomegalovirus (CMV) retinitis. The presence of microangiopathy was not related to TB. In Malawian patients with TB presenting acutely with fever, choroidal granulomas were found in 2.8%, and were concurrent with mycobacteraemia and AIDS. Ophthalmoscopy was not a useful aid in the diagnosis of mycobacteraemia. CMV retinitis is rarely seen in African AIDS patients. This may be due to mortality early in the disease course, or differences in race, HIV sub-type or co-morbidity.

Multiple human herpesvirus-8 infection.

In Malawian patients with Kaposi sarcoma (KS) and their relatives, we investigated nucleotide-sequence variation in human herpesvirus-8 (HHV-8) subgenomic DNA, amplified from oral and blood samples by use of polymerase chain reaction. Twenty-four people had amplifiable HHV-8 DNA in >1 sample; 9 (38%) were seropositive for human immunodeficiency virus type 1, 21 (88%) were anti-HHV-8-seropositive, and 7 (29%) had KS. Sequence variation was sought in 3 loci of the HHV-8 genome: the internal repeat domain of open-reading frame (ORF) 73, the KS330 segment of ORF 26, and variable region 1 of ORF K1. Significant intraperson/intersample and intrasample sequence polymorphisms were observed in 14 people (60%). For 3 patients with KS, intraperson genotypic differences, arising from nucleotide sequence variations in ORFs 26 and K1, were found in blood and oral samples. For 2 other patients with KS and for 9 people without KS, intraperson genotypic and subgenotypic differences, originating predominantly from ORF K1, were found in oral samples; for the 2 patients with KS and for 4 individuals without KS, intrasample carriage of distinct ORF K1 sequences also were discernible. Our findings imply HHV-8 superinfection.

Pathophysiology of cerebral malaria: role of host cells in the modulation of cytoadhesion.

Cerebral malaria (CM), one of the most serious complications of Plasmodium falciparum infection, is characterized by the sequestration of infected erythrocytes (IEs) in cerebral microvascular beds. The precise mechanisms involved in the onset of neuropathology remain unknown, but parasite sequestration in the brain, metabolic disturbances, and host immune responses all play a role. Studies in a murine model of CM showed a potential role for host cells, especially platelets, in the pathogenesis of CM. Indeed, urokinase plasminogen activator receptor (uPAR; CD87) deficiency attenuates the severity of CM, most likely by its important role in platelet kinetics and trapping. These results led us to evaluate whether platelets have a role in the human disease. By immunostaining of brain samples from Malawian patients, we determined that the surface of platelet accumulation and the proportion of vessels filled with platelets were significantly higher in patients who died of CM than in those who died of other causes. We then investigated the role of platelets in IE cytoadhesion in vitro, using CD36-binding IE (IECD36) and CD36-deficient (CD36DEF) brain microvascular endothelial cells (ECs). Coincubation studies indicated that platelets can induce strong IECD36 binding to CD36DEF ECs and, conversely, can hide constitutively expressed falciparum receptors such as chondroitin sulfate A. Thus, platelets may provide an adhesion receptor to microvascular beds originally devoid of it. This novel mechanism of cytoadhesion may reorient the sequestration of different parasite phenotypes and play an important role in the pathogenesis of severe malaria.

Demographic and immune correlates of human herpesvirus 8 seropositivity in Malawi, Africa

Background: In the USA, human herpesvirus 8 (HHV-8) is associated with Kaposi's sarcoma (KS) and HIV infection. We examined HHV 8 seroprevalence in a Malawian cohort, and assessed its relationship with HIV, KS, demographic characteristics, and immune findings. Methods: In 1997 and 1998, blood samples were obtained from 272 hospitalized Malawian patients, for whom demographic information was obtained, and 24 healthy volunteers without demographic data. We used enzyme immunoassays to assess seroprevalence and antibody titers to peptide antigens derived from HHV-8 K8.1 and ORF65-encoded proteins. Intracellular cytokines and cell surface antigens were assessed with four-color flow cytometry. Data were analyzed using non-parametric univariate and regression analytic techniques. Results: The rates of HHV-8 seroprevalence to either or both HHV-8 peptides were 67% for the patients and 54% for the healthy volunteers. Seroprevalence increased with patients' age (P<0.001) but was not associated with HIV status, percentage of lymphocytes expressing CD4, or KS (n=10). Seropositive females had lower antibody titers to both peptides than did males (medians: 455 versus 1361 for K8.1, P<0.001; and 268 versus 405 for ORF65, P=0.044). For the healthy volunteers, the percentage of CD8+ cells producing IFN-y after stimulation was significantly lower in ORF65-specific antibody-positive persons (medians: 24% versus 57%, P=0.008). Conclusions: In Malawi, HHV8 is endemic and is not associated with HIV infection or HIV severity. Seroprevalence rates increase in childhood, and, most steeply in adolescence. Titers are higher in seropositive males than in sero-positive females. The immune effects of HHV-8 in healthy adults are consistent with chronic inhibition of type 1 cytotoxic T -cell responsiveness, independent of HIV status.

Age-related differences in cell-specific cytokine production by acutely ill Malawian patients.

Age-related changes in human cell-specific cytokine responses to acute illness have not been well examined. We therefore evaluated age-related differences in T, B and natural killer (NK) peripheral blood lymphocyte cytokine responses of 309 acutely ill hospitalized people in Malawi, Africa, < 1 month-61 years of age. We used four-colour flow cytometry and performed Wilcoxon rank sum and Kruskal-Wallis tests, Pearson (rp) and Spearman (rs) correlations, and linear and logistic regression analyses to control for human immunodeficiency virus infection (HIV) status, the percentages of lymphocytes expressing CD4, and the nature of the acute infection. The percentages of CD8- and CD8+ T cells producing induced IL-8 decreased with age (rs = -0.44 and -0.53). The percentages of T cells producing TNF-alpha were higher, and the percentages producing IL-10 were lower, in those > or =13 than those < 13 years old (medians: 17.7 versus 10.5 and 1.4 versus 3.0, respectively). The percentages of CD8- T cells producing IFN-gamma were higher and stable in those > or =1 year old compared to infants (medians: 23.5 versus 10.4); the percentages of NK producing IFN-gamma were higher post-infancy and then declined to relatively low levels with increasing age. The percentages of T cells producing IL-2 were highest in those 5- <31 years old (median 5.6) and lowest in those > or =31 years old (median 1.9). The ratios of the percentages of T cells producing IL-4 to those producing IL-8 and to those producing IL-10 both increased with age. These data suggest that innate immunity, represented by NK IFN-gamma production, dominates in early life. A number of shifts occur after infancy and before adolescence, including a proinflammatory shift from IL-8 to TNF-gamma and a type 2 shift from IL-10 to IL-4 dominance. These findings suggest distinct age-related differences in the human response to acute illness and may be useful in directing future efforts at immunomodulatory therapies.

Plantar and digital dermatoglyphic patterns in Malawian patients with diabetes, hypertension and diabetes with hypertension

In this study we examined the dermatoglyphic features of 99 Malawian patients with diabetes,hypertension and diabetes with hypertension, attending the outpatient clinic at Lilongwe and QueenElizabeth Central Hospitals. Plantar and digital pattern types were determined and their variabilitywas calculated using Dankmeijer’s (DI) and pattern intensity (PII) indices. The results showed thatthe soles of all patients had more loops than arches and more arches than whorls, which wererestricted to the distal zones. In hypertension, whorls were found in zones I, II and III whereas inpatients with diabetes and hypertension, the whorls were seen in zones I, III and IV. In digits themost predominant ridge pattern were arches in all patients, followed by loops and whorls were absent.In the first digit, diabetic patients had no arches but women hypertensives showed arches. In patientswith diabetes and hypertension, arches were present in both sexes but in men it was confined to theright foot. Loops were only found in the first digit in all patients. The frequency of loops was highestin diabetic patients, high in diabetics with hypertension and least in patients with hypertension alone.Using PII and DI indices, there were significant differences between patients and healthy Malawiansubjects (P < 0.001), and between the sexes (P < 0.01), but PII did not show significant differenceswith sex (P > 0.5) in hypertensives. Diabetic patients had the lowest mean DI when compared to theother groups. In conclusion, this study has demonstrated plantar and digital dermatoglyphicdifferences between patients and healthy Malawians, differences that were exhibited more in digitalthan in plantar ridge patterns. The disease conditions were indicated better by whorls intopographical zones IV of the sole and by the presence and frequency of loops in the first digit.

African Burkitt's lymphoma: a new perspective

High titres of antibody to Epstein-Barr virus (EBV) late genes identify individuals at risk of developing endemic Burkitt's lymphoma (eBL). Viral lytic cycle early and intermediate-early gene expression in BL is associated with a favourable tumour response to chemotherapy. Our study investigated whether serological data identifying antibody expression to zta, a viral function that activates lytic-cycle gene expression, correlate with expression of its gene in tumours, and could have prognostic value. Studies on 10 Malawian patients, with presumed BL on clinical grounds, showed good correlations, suggesting that serum antibody responses might predict treatment responsiveness. The results with 1 patient were particularly striking. When admitted in January 1998, prognosis was poor as he was unable to walk, and had tumour cells, characteristic of stage IV disease, in his bone marrow. Laboratory investigations showed particularly high levels both of serum zta antibodies and of gene expression in his tumour. Follow-up confirmed him alive 6 months after hospital discharge. Among the EBV-positive cases, 2 were ultimately diagnosed as rhabdomyosarcoma, a tumour not previously associated with this virus. The findings from this small study, if confirmed, should have value for future BL management in resource-poor parts of the world.

Carcinoma of the conjunctiva and HIV infection in Uganda and Malawi.

To investigate the association of human immunodeficiency virus (HIV) infection and carcinoma of the conjunctiva in Africa, and the role of human papilloma virus type 16 (HPV-16). Patients in Uganda and Malawi presenting to eye clinics with lesions suspicious of carcinoma were studied. Pathological confirmation of eye lesions was sought. HIV testing of patients who were biopsied and, in Uganda, of matched case control subjects was carried out as was testing of a sample of fixed biopsies for HPV-16 by polymerase chain reaction (PCR). The HIV-1 serology, histopathology of conjunctival biopsies (conjunctival intraepithelial neoplasia (CIN), invasive carcinoma, other lesions), and prevalence of HPV-16 infection were determined. Of Ugandan patients, 27/38 (71%) with carcinoma (27 invasive carcinoma, 11, CIN) were HIV positive compared with 12/76 (16%) of controls (odds ratio 13, 95% confidence interval 5-38). The calculated population aetiological fraction of carcinoma associated with HIV was 66%. Of 32 Malawian patients (20 invasive carcinoma, 12 CIN), 25/29 tested (86%) were HIV positive. HPV-16 infection was found in 7/20 (35%) of carcinoma samples, 0/9 pingueculae, and 2/6 conjunctivitis samples. HIV infection is strongly associated with an apparent increase in the incidence of conjunctival carcinoma in Africa. While ultraviolet light is probably the prime risk factor and HPV-16 is implicated in a proportion of cases, the interactions of ultraviolet light, HIV, HPVs, and other factors are unclear in the pathogenesis of carcinoma. The disease represents another model of multifactorial epithelial carcinogenesis.

Retinal findings in Malawian patients with AIDS.

The purpose of this study was to determine the prevalence of retinal disease in a group of patients with AIDS in Malawi. Indirect ophthalmoscopy was performed on 99 patients with AIDS who were admitted consecutively to a medical ward at a central hospital. Necrotising retinitis was present in one eye of one patient examined. Non-infectious retinopathy was present in 13%. Retinitis is less common in AIDS patients from Africa compared with those from developed countries; it is believed that most patients die before acquiring it. Non-infectious retinopathy may also be less common.

Clinical Diagnosis of Tamponade in Malawi

A consecutive series of 25 Malawian patients with tamponade secondary to tuberculosis were compared to 25 patients with congestive cardiac failure, without pericardial effusion in a retrospective study. More patients with tamponade had an impalpable apex beat (21/1), pulsus paradoxus (13/0), soft heart sounds (13/2), paradoxical rise in jugular venous pressure (6/0), and fewer had a murmur (1/14). All these results are significant (p less than 0.05) by the chi 2 test with Yates' correction. The presence of two or more of these discriminating physical signs has a positive predictive value of 75 per cent, and a negative predictive value of 99.5 per cent for the diagnosis of tamponade. Clinical diagnosis of tamponade by primary health care personnel in Malawi should be possible, and lead to earlier treatment.

Lumbar puncture in Malawian patients with altered consciousness.

Lumbar puncture in Malawian patients with altered consciousness.

A geographical study of antibodies to membrane antigens of HSV-2-infected cells and HSV-2-specific antibodies in patients with cervical cancer.

Sera was obtained from patients with squamous carcinoma of the cervix from Great Britain (29), Sri Lanka (32), Malawi (27), and Sudan (27), and from controls from all countries except Sudan. Controls were matched for ethnic origin, age and social class. Sera were tested by indirect immunofluorescence (IF) for IgG and IgA antibodies to membrane antigens (MA) and IgA antibodies to VCA of HSV-2 infected cells. Compared with controls, IgA antibodies to MA (IgA anti-MA) were detected more frequently and at higher titres in all groups of patients. However, there was no significant difference in prevalence of these antibodies at titres greater than or equal to 1:4 between Malawian patients and controls, although a significantly higher proportion of patients had IgA anti-MA titres of greater than or equal to 1:16. In contrast, IgA anti-VCA did not distinguish patients from controls. More than 90% of both patients and controls from all countries had IgG antibodies to MA (IgA anti-MA). Malawian patients had a significantly higher geometric mean titre (GMT) of IgG anti-MA than controls and both patients and controls had significantly higher GMTs than their counterparts from other countries. The variation between herpes IgG anti-MA titres in subjects from different countries did not reflect differences in serum immunoglobulin levels and similar variations in titre were not seen in rubella and measles HAI titres. Among the patients there was a geographical variation in the prevalence of HSV-2 specific antibodies detected by ELISA, which varied from 52% among British and Sudanese patients to 73% among Malawian patients. Even when adjustment was made for possible false negative results, there were between 10 and 31% patients without HSV-2-specific antibodies, although only 2 of 103 (1.9%) patients had neither HSV-1 nor HSV-2 antibodies. The association of HSV-2 with cervical carcinoma appeared to vary with age.

Venereal syphilis in tropical Africa.

A steady decline in the incidence of positive results to the Kahn test is reported in Malawian patients during the period 1968-75. Other studies have shown that the incidence of early and late syphilis in sub-Saharan Africa has dropped considerably over the past few decades. The number of reported cases of early syphilis in certain urban areas, however, appears to be high. It is suggested that the downward trend in the incidence of syphilis in Africa is related to the increased and often indiscriminate use of penicillin.