Prevalence of hypertension and correlates among adults of 45-60 years in a rural area of Tamil Nadu.

Hypertension is one of the major causes of cardiovascular morbidity and mortality. Community based studies in the rural areas of Tamil Nadu on the prevalence of hypertension and its associated risk factors are scarce. A cross-sectional study was undertaken among a sample of 406 individuals (45-60 years) selected by the standard 30 cluster systematic random sampling technique to find out prevalence of hypertension and its associated risk factors in a rural area of Tamil Nadu. Chi-square test and multiple logistic regression were employed using SPSS package. The overall prevalence of hypertension was 33% and higher among sedentary type (41%). In bivariate analysis many of the independent variables correlated with hypertension, but in multivariate analysis, only body-mass index, family history and age remained significant.

First time detection of Japanese encephalitis virus antigen in dry and unpreserved mosquito Culex tritaeniorhynchus Giles, 1901, from Karnal district of Haryana state of India.

Japanese encephalitis virus (JEV) antigen has been detected by antigen capture enzyme linked immunosorbentassay (ELISA) in dry specimens of the mosquito Culex tritaeniorhynchus Giles, 1901, collected from Karnal district of Haryana state in northern India. These mosquitoes were stored in dry condition for 20 months, at room temperature, before processing. The procedure of detecting JEV infection in long time stored, dry vector mosquitoes, has important application in the surveillance of Japanese encephalitis.

T cell reactivity of defined peptides from a major Plasmodium falciparum vaccine candidate: the Pf155/RESA antigen.

Several immunodominant B-cell epitopes of the P. falciparum antigen blood stage Pf155/RESA, a major vaccine candidate antigen, are located in the molecular regions containing amino acid repeats. We started to map Pf155/RESA for T cell reactive epitopes. For this purpose, short synthetic peptides corresponding to the 3'- and 5' repeat regions of the molecule as well as to non-repeated sequences outside these regions were prepared. T cells from P. falciparum primed donors from two highly endemic areas of Africa were tested for their responsiveness to the peptides by thymidine incorporation and/or interferon gamma (IFN-gamma) release. There was a considerable variation in the response to the different peptides. However, the strongest and most frequent responses were seen with a few peptides from the 3'- and 5'-repeat regions. Thus, the immunodominant B cell epitope regions of Pf155/RESA, contain several T cell epitopes. Since the repeat regions are known to be conserved in different P. falciparum strains, the T cell epitopes reported here may be suitable constituents of a P. falciparum subunit vaccine.

Characterization of regulatory T cell responses to defined immunodominant T cell epitopes of the Plasmodium falciparum antigen Pf155/RESA.

Several immunodominant B and T cell epitopes of the P. falciparum blood stage antigen Pf155/RESA, a vaccine candidate, are located in the central (5') and C-terminal (3') invariant repeat regions of the molecule. Here we have attempted to functionally analyze human T cell responses to some of the T cell epitopes. For this purpose short synthetic peptides corresponding to these epitopes were used to study the induction of in vitro expression of IL-4 mRNA, IFN-gamma secretion, proliferation and B cell help for antibody production. In individual malaria immune donors these different T cell activities were not correlated. The findings emphasize the importance of examining multiple parameters of T cell activation when estimating the total proportion of individuals responding to a defined antigen. IL-4 mRNA was expressed in activated T cells of donors who had elevated serum concentrations of antibodies to the peptide used for T cell activation. These results suggest the involvement of IL-4 producing T helper cells in the induction of Pf155/RESA specific antibody production in individuals in which immunity has been induced by natural infection. Taken together, these findings also suggest that functionally distinct CD4+ T cells occur in humans similarly to what has been described in mice. In further experiments, we have also attempted to establish MHC class II restriction of the immune response to these epitopes at the level of the donor populations. When studying monozygotic twins, antibody responses to Pf155/RESA derived peptides and some of the T cell responses could be paired within the twin pairs, indicating a genetic regulation of their B cell responses. Whether or not this regulation reflects MHC class II restriction, or other factors needs to be elucidated.

Immunogenicity of the nonrepetitive regions of the circumsporozoite protein of Plasmodium knowlesi.

The circumsporozoite antigen (CS) of the simian malarial parasite Plasmodium knowlesi consists of tandemly repeated immunodominant peptide units that are variable and may play a role in evading the immune system. To study the immunogenicity of this antigen in the absence of the immunodominant repeats, the entire nonrepetitive region of the antigen was expressed in Escherichia coli as two fusion proteins with glutathione-S-transferase (GST) representing the amino terminal (GST-CSN) and the carboxy terminal domains (GST-CSC) of the CS antigen. The immunogenicity of these fusion proteins was studied in rabbits and different strains of mice. Antibody raised against both the CSN and CSC domains in both rabbits and every strain of mice recognized the native protein, as detected by immunofluorescence assay (IFA) using P. knowlesi sporozoites. A positive IFA reaction was also obtained with P. vivax sporozoites using antisera raised against the CSC domain. High titer antisera were raised in rabbits against both the domains, whereas mice showed comparatively low titers. On Western blots, mice showed specific response against the CSC domain. In both rabbits and mice, significant titers of antibodies were raised against region II, which has been shown to be the putative sporozoite binding site for hepatocytes in the case of P. falciparum.

An epidemiological study of humoral and cell-mediated immune response to the Plasmodium falciparum antigen PF155/RESA in adult Liberians.

We investigated the seroreactivity and T cell reactivity against the Plasmodium falciparum antigen Pf155/RESA, different oligopeptides from the 3' and 5' repeat regions of the Pf155/RESA antigen, and crude Plasmodium falciparum antigens in 164 adult Liberians. We compared 2 long-term residential groups with high and low exposure to malaria. The seropositive rate to the peptides was significantly higher with increased exposure. There was no significant difference in response rates to the Pf155/RESA. This may indicate the level of persistent T cell memory in previously primed donors. The seropositive rates to 3 Pf155/RESA peptides and the rates measured by either 3H-thymidine incorporation or IFN-gamma release after stimulation with Pf155/RESA and the peptides were all lower in parasite positive individuals. Even low grade, asymptomatic parasitemia can impair the T cell response in vitro. The lower antibody response in parasite positive subjects may be explained by either antibody consumption or lower protection against malaria parasitemia in subjects with low concentrations of antibodies against the Pf155/RESA antigen.

T cell immunity in malaria.

Immunity to malaria is a complex process. In malaria endemic areas, sustained clinical immunity develops gradually. Protective immunity to malaria comprises both antibody-dependent and antibody-independent effector mechanisms. Although, the relative roles of B and T cells differ in different malaria systems and different stages of the parasite, T cells are essential for the induction and maintenance of immunity against malaria. T cell-derived soluble factors are believed to be important mediators of cellular effector mechanisms. The efficacy of an antigen as a malaria vaccine depends to a great extent on the T cell recognition sites and the nature of the responses induced by these determinants. In order to select suitable epitopes in an immunogen, an understanding of the definition of antigenic sites recognized by T cells and characterization of the nature of the T cell responses induced in malaria endemic populations is important.

A modified immunofluorescence assay for detection of Japanese encephalitis virus-infected cells.

Japanese encephalitis virus (JEV) infections are currently detected by indirect immunofluorescence (IF) assay using virus-specific antibodies on acetone-fixed smears. On few occasions, the acetone treatment was reported to damage certain epitopes on JE virus (JEV) glycoprotein. Here, we have made an attempt to adopt quick paraformaldehyde fixation followed by a short detergent treatment of cells in suspension for identification of JEV-infected brain cells of laboratory-reared Toxorhynchitis splendens mosquito larvae using virus-specific antibodies. JEV-positive cells could be scored by the presence of a well defined intracellular immunofluorescence staining against unstained uninfected antibody-treated cells. The advantage of this assay is that stained cell suspensions can be stored for up to 4 weeks, allowing analysis at convenience. Thus, the modified IF assay can be employed as an additional/alternate technique to standard IF assay for detection of JEV in cells and also to screen hybridoma cell lines for anti-JEV antibody production.

Humoral immune responses to the Plasmodium falciparum antigen Pf155 RESA in adults with differential clinical conditions from an Indian zone where malaria is endemic.

Ring infected erythrocyte surface antigen of Plasmodium falciparum (Pf155/RESA) has been considered as a vaccine candidate. However, the relative immunogenicity of this antigen has not been studied in Indian populations. Pf155/RESA was investigated for its immunogenicity by studying humoral immune responses against Pf155/RESA and Pf155/RESA derived peptides (P1, P2 representing immunodominant epitopes from the 3' and P3 from the 5' repeat regions) by erythrocyte membrane immunofluorescence (EMIF) assay and by enzyme linked immunosorbent assay (ELISA) in P. falciparum primed donors living in hyperendemic malarious areas (Orissa State, India) where P. falciparum infections are highly prevalent. Subjects of different clinical status namely acute (A), clinically immune (CI) and acute with history of repeated P. falciparum infections (R) were included in this study. All the donors were seropositive against the crude antigen. There was considerable variation in the responses among the donors. While humoral responses in the plasmas against the P2 peptide (EENV)4 were significantly higher in magnitude and in frequency in the CI donors than in the A donors, no positive response was seen in the R donors. The responses to the peptides P1 (EENVEHDA)2 and P3 (DDEHVEEPTVA)2 were poor both in the A and in the CI groups. Whereas, most of the R donors were seropositive against the P3. The present results indicate that Pf155/RESA contains B cell epitopes which were recognized differently by the immune system of individuals living in malaria-hyperendemic areas of India who have been primed by natural infection. Our studies also suggest that in order to investigate the possible functional role of a given antigen, study of immune responses against the antigen in donors of different clinical status may be useful.

Cross-resistance to Bacillus sphaericus strains in Culex quinquefasciatus resistant to B. sphaericus 1593M.

Bacillus sphaericus 1593M resistant larvae of Culex quinquefasciatus were reared in the laboratory since 1995. Resistance in the larvae was monitored by subjecting selection pressure using B. sphaericus 1593M at every generation. Bioassays were conducted with different strains of B. sphaericus (Bs 2297, Bs 2362 and Bs IAB 59) and confirmed cross-resistance in the present study. The level ranged between 27.3 to 18.2 fold in comparison with susceptible larvae. But Bacillus thuringiensis var israelensis strains (Bti PG14 and Bti 426) did not show any cross-resistance in the larvae and it emphasized a need to study the mode of action of B. sphaericus toxin that induces cross-resistance in the larval strain.

Immune responses to chloroquine--sensitive and resistant populations of Plasmodium berghei in mice.

In order to elucidate the role of the host as a factor in the spread of chloroquine resistance, a study of the host's immune responses in chloroquine resistant (cqr) and chloroquine sensitive (cqs) Plasmodial infections is essential. Course of the infection and the nature of immune responses in mice infected with chloroquine resistant (R) and chloroquine sensitive (S) strains of Plasmodium berghei were compared. Crude parasite antigen activated T cells from both the groups of mice (R and S) and the parasite specific antibodies were detected in the sera of both the groups. The differences in immune responses between the control (uninfected) and infected mice were found to be significant. However, humoral and in vitro cellular responses obtained with T cells from chloroquine resistant P. berghei primed mice was lower in comparison with the responses obtained with T cells from the sensitive infections. Our studies therefore suggest that immunosuppression to parasite antigen is seen in mice primed with chloroquine resistant P. berghei, which may play a role in the development of resistance.

The 2001 dengue epidemic in Chennai.

UNLABELLED: Dengue is emerging as a serious public health problem in Tamil Nadu. The present surveillance system is unlikely to generate proper information on the epidemiology of the disease, which is essential for planning and development of relevant control/preventive measures against dengue. OBJECTIVE: Between November 2001 and January 2002, a descriptive study was undertaken on children with clinical dengue attending Kanchi Kamakoti Child Trust Hospital (KKCTH, a major private referral pediatric hospital in Tamil Nadu, India) to define the magnitude of dengue burden, the natural history of this disease in terms of clinical presentation, and outcome of the infections in hospitalized children (< 15) with clinical dengue. METHODS: The sera collected from patients analyzed for dengue specific IgM and IgG antibodies by IgM, IgG antibody capture enzyme linked immunosorbent assay (ELISA) using alternatively two commercial kits. World Health Organization clinical case definition was adopted to categorize the dengue confirmed children. RESULTS: Dengue was diagnosed in 74.5% (143) of the 192 hospitalized children with clinical dengue. A considerable proportion (20%) of the total dengue infections were constituted by infants less than 1 yr of age. DF [dengue fever], DHF [dengue hemorrhagic fever] and DSS [dengue shock syndrome] were diagnosed in 65%, 11.2% and 23.8% of 143 dengue confirmed patients respectively. Though severe dengue (DSS + DHF) was present in 35% of the patients, the mortality rate was low during the study period due to timely diagnosis and clinical management of the patients. CONCLUSION: In developing countries like India, building of laboratory capacity for diagnosis and combat-mode ready preparedness for the management of dengue cases in emergency situation may reduce dengue-related mortality. This can be achieved in a wider scale through an integrated approach through the community, professionals and the public health departments.