Isolation and Evaluation of Diagnostic Value of Two Major Secreted Proteins of Mycobacterium Tuberculosis

Two secreted antigens of Mycobacterium tuberculosis, namely the antigen 85 complex (30/31) and 38kDa antigens, were purified from the whole culture filtrate by using two dimensional preparative electrophoresis and anion exchange chromatography, respectively. Individual components of the antigen 85 complex namely, antigen 85A, 85B and 85C, were separated using hydrophobic interaction chromatography. The humoral antibody activity to these antigens in sputum positive cases of active pulmonary tuberculosis and normal healthy volunteers was determined by enzyme linked immunosorbent assay (ELISA) and immunoblot. Recombinant 38kDa and antigen 6 were used as reference antigens for the assay. None of the healthy volunteers reacted with the 38kDa antigen, while 52% of the TB sera reacted with it. Of the three components of the antigen 85 complex, 85B gave the highest positivity of 40 per cent. The results of combination of 38kDa with antigen 6 offered better results with 76% positivity

Age-related changes in blood lymphocyte subsets of south Indian children

Background. Enumeration of lymphocyte subsets has been widely used for the diagnosis and monitoring of several haematological and immunological disorders. Various studies have demonstrated age, sex and racial differences in lymphocyte subset expression. Reference values are not available for Indian children and there is a need for this information to replace commonly used, but inappropriate, adult lymphocyte subset ranges. Methods. One hundred thirty-eight healthy children be tween 3 and 15 years of age, attending a local government school in Chennai, South India were included in the study. Haemoglobin levels, and total and differential cell counts were determined using an automated counter and lymphocyte subsets were analysed by flowcytornetry. Results. The mean (SD) absolute lymphocyte count declined with age from 4338 (1031) at 3 years to reach a plateau of 3096 (914) at 11-13 years (p < 0.05). A significant decline was also observed in the absolute numbers of CD3+, CD4+, CD8+ and CD19 + cells. However, the percentage values of CD3+, CD4+, CD8+, CD16/56+ cells and the CD4/CD8 ratio remained fairly stable across the age range. Conclusion. Our data would prove useful in interpreting disease-related changes in lymphocyte subsets in Indian children of different age groups. Age-related decrease in the absolute lymphocyte count as well as numbers of CD4 and CD8 cells was found to occur between the ages of 3 and 11 years. A normogram relating age to CD4 count has been developed

Cell-mediated immunity in chyluria

Cellular immune response to mitogens phytohemaggluthin (PHA) and poke weed mitogen (PWM) was assessed in 13 patients with chyluria and 32 healthy controls. The mean stimulation Index of the patient group was significantly lower than the control group. The degree of depression was neither related to the duration of excretion of chyle nor to the microfilaraemic status

Hepatic Vascular Variants in Hereditary Haemorrhagic Telangiectasia: Imaging findings

Hereditary Haemorrhagic Telangiectasia (HHT) is an autosomal dominant disorder characterized by vascular dysplasia. Hepatic Vascular Malformations (VMs) range from small telangiectases to significant vascular shunting. Here we report two cases of HHT. Case 1 had diffuse ectasia of the hepatic artery along its intrahepatic and extrahepatic course with a hepatic arterial aneurysm. Case 2 presented with ileal and hepatic telangiectases. Knowledge of these vascular variants is indispensable for clinicians and radiologists in aiding diagnosis and surgical and interventional management. Keywords: Vascular Malformations, HHT, Arteriovenous Malformation, Ileal Telangiectasis

HLA-DR phenotypes and IgG, IgA and IgM antibody responses to Mycobacterium tuberculosis culture filtrate and 30 kDa antigens in pulmonary tuberculosis

The role of HLA-DR genetic make-up on the IgG, IgA and IgM antibody response to Mycobacterium tuberculosis culture filtrate and 30 kDa antigens was studied in pulmonary tuberculosis. The study was carried out in HLA-DR typed active pulmonary tuberculosis (ATB) patients (n = 37), inactive (cured) pulmonary tuberculosis (ITB) patients (n = 79) and normal healthy subjects (NHS; n = 46). In ATB and ITB (cured) patients, IgG antibody (optical density at 490 nm for 1 : 3200 dilution) as measured by enzyme-linked immunosorbent assay was the predominant one than IgA and IgM antibodies. Increased IgG antibody titre to culture filtrate (P = 0.03) and decreased titre to 30 kDa antigen were observed with HLA-DR1-positive ATB patients than non-DR1 (ATB) patients. Moreover, HLA-DR4- and HLA-DR6-positive ATB patients showed trends toward an increased IgG antibody response to 30 kDa antigen than HLA-DR4- and HLADR6- negative (ATB) patients respectively. Significantly increased IgA antibody to 30 kDa antigen was observed with HLA-DR1-positive ATB patients than non-DR1 patients (P = 0.03). The study suggests that multiple HLA-DR molecules may regulate the IgG and IgA antibody responses to various proteins of M. tuberculosis. Moreover, HLA-DR phenotypes and increased IgG and IgA antibody titres may be useful to differentiate M. tuberculosis-infected subjects from normal subjects and cured patients with the same HLA-DR phenotypes or genetic make-up

Yield of QuantiFERON-TB gold in tube assay and tuberculin skin test in healthy persons from a tuberculosis endemic population

Abstract. We assessed the yield of the QuantiFERON-TB Gold in tube test (QFT-GIT) and the tuberculin skin test (TST) in healthy persons from our general population, where tuberculosis is endemic and Bacillus Calmette-Gu´erin is mandatory. The yield of QFT-GIT (2%) and TST (1%) were very low among children, where the test was positive in 42% and 24% of adults, respectively. Our study results show that QFT-GIT and TST are not influenced by prior Bacillus Calmette-Gu´erin vaccination. Applying the reduced cut-off point of 0.13 IU/mL is adequate in a pediatric population. Moreover, both the TST and the QFT-GIT demonstrated a low yield in the pediatric population, which suggests that either TST or QFT-GIT alone or in combination can be used to diagnose Mycobacterium tuberculosis infection in our setting

Cellular immune response to Mycobacterium tuberculosis-speciWc antigen culture Wltrate protein-10 in south India

The Mycobacterium tuberculosis (M. tuberculosis)- speciWc culture Wltrate protein-10 (CFP-10) is highly recognized by M. tuberculosis infected subjects. In the present study, the proliferative response and IFN-� secretion was found for C-terminal peptides of the protein (Cfp651–70, Cfp761–80, Cfp871–90, and Cfp981–100). The alleles HLA DRB1 *04 and HLA DRB1 *10 recognized the C-terminal peptides Cfp7, Cfp8, and Cfp9 in HHC. Cfp6 was predominantly recognized by the alleles HLA DRB1 *03 and HLA DRB1 *15 by PTB. The minimal nonameric epitopes from the C-terminal region were CFP-1056–64 and CFP-1076–84. These two peptides deserve attention for inclusion in a vaccine against tuberculosis in this region

Immune response to Mycobacterium tuberculosis specific antigen ESAT-6 among south Indians

The 6-kDa early secreted antigenic target (ESAT-6) is a T-cell antigen recognized by individuals infected with Mycobacterium tuberculosis. The aim of the study was to identify ‘‘protective epitopes’’ of ESAT-6 protein in the south Indian population. Proliferative and Interferon gamma (IFN-g) responses to ESAT-6 peptides were studied by flow cytometry and Enzyme linked immunosorbent assay (ELISA). Healthy household contacts (HHC) recognized Esp1 (10/17) and Esp6 (9/17) peptides. Among pulmonary tuberculosis patients (PTB), Esp1 (3/11) and Esp6 (5/11) were recognized. Maximal response (7/10) was found for Esp1 and Esp8 in treated patients (TR). Median values for the responding subjects gave the following results: Esp1 (76 pg/ml), Esp6 (64 pg/ml), induced IFN-g production in HHC; PTB gave low IFN-g responses for the peptides. TR responded to the peptides Esp1 (141 pg/ml), Esp8 (102 pg/ml). The proliferation of CD4 cells was similar in both PTB and TR for all peptides; but HHC showed an increase for Esp1 (p < 0.05) and Esp6 (p < 0.01). Esp1 (amino acids aa 1–20) and Esp6 (aa 51–70) were the immunogenic peptides recognized by the alleles HLA DRB1*04 and HLA DRB1*10 among HHC. But the association of the alleles with ESAT-6 peptide presentation needs to be confirmed in a large cohort of subjects. We speculate that ESAT-6 can be used along with other immune-eliciting proteins for vaccine design strategies in south Indian population

Humoral immune response to filarial antigens in chyluria

Humoral immune parameters like total immunoglobulins and specific antibody levels in serum were studied in filarial chyluria patients. Mean serum IgG was significantly reduced in this group compared to normal controls, while IgA and IgM levels remained comparable to controls. Anti-filarial antibody titre as measured by enzyme-linked immunosorbent assay also was significantly reduced. However, the total and specific IgE antibody titre was similar to that of controls. Specific IgE contents of the patients’ sera could be related to their microfilaraemic status

Expert United Kingdom consensus on the preservation of joint health in people with moderate and severe haemophilia A: A modified Delphi panel

Aim: For people with haemophilia A (PwHA), bleeding in the joints leads to joint damage and haemophilia-related arthropathy, impacting range of motion and life expectancy. Existing guidelines for managing haemophilia A support healthcare professionals (HCPs) and PwHA in their efforts to preserve joint health. However, such guidance should be reviewed, considering emerging evidence and consensus as presented in this manuscript. Methods: Fifteen HCPs experienced in the management of PwHA in the UK participated in a three-round Delphi panel. Consensus was defined at ≥70% of panellists agreeing or disagreeing for Likert-scale questions, and ≥70% selecting the same option for multiple- or single-choice questions. Questions not reaching consensus were revised for the next round. Results: 26.8% (11/41), 44.8% (13/29) and 93.3% (14/15) of statements reached consensus in Rounds 1, 2 and 3, respectively. HCPs agreed that prophylaxis should be offered to patients with a baseline factor VIII (FVIII) level of ≤5 IU/dL and that, where there is no treatment burden, the aim of prophylaxis should be to achieve a trough FVIII level ≥15 IU/dL and maintain a longer period with FVIII levels of ≥20-30 IU/dL to provide better bleed protection. The aspirational goal for PwHA is to prevent all joint bleeds, which may be achieved by maintaining normalised (50-150 IU/dL) FVIII levels. Conclusion: The panel of experts were largely aligned on approaches to preserving joint health in PwHA, and this consensus may help guide HCPs