High microbial translocation limits gut immune recovery during short-term HAART in the area with high prevalence of foodborne infection.

BACKGROUND: Individuals residing in areas with high prevalence of foodborne infection could have a higher risk of gut microbial translocation which may affect monocyte activation, gut immune recovery and intestinal epithelial cell damage. We aimed to measure alterations in microbial translocation, monocyte activation, gut immune recovery, and intestinal epithelial cell damage in HAART treated individuals. METHODS: A prospective, single-arm, longitudinal, cohort study was conducted among antiretroviral naïve HIV-1 infected Thai participants. All participants were in chronic stage of HIV-1 infection before starting HAART. Data and samples were collected prior to initiation of HAART and then after 24 and 48 weeks of HAART. Plasma biomarkers for microbial translocation (16S rDNA and LBP), monocyte activation (sCD14) and intestinal epithelial cell damage (I-FABP) were evaluated. We measured circulating gut-homing CD4+ T cells and circulating gut-homing Th17 cells to assess recoveries of gut immunity and gut immunity to microbial pathogens. RESULTS: The kinetic studies showed no reduction in the levels of plasma 16S rDNA, sCD14 or I-FABP, significant decrease of plasma LBP level, and slow but significant increases in the frequencies of circulating gut-homing CD4+ T cells and circulating gut-homing Th17 cells during 48 weeks of HAART. Dividing participants into low and high microbial translocation (low and high MT) groups at baseline, both groups showed persistent plasma levels of 16S rDNA, sCD14 and I-FABP, and significantly decreased plasma level of LBP. The low MT group had significantly increased frequencies of circulating gut-homing CD4+ T cells and circulating gut-homing Th17 cells during 48 weeks of HAART but this was not observed in the high MT group. CONCLUSIONS: We demonstrated persistent high microbial translocation, monocyte activation and intestinal epithelial cell damage with slow gut immune recovery during successful short-term HAART. Additionally, gut immune recovery was apparently limited by high microbial translocation. Our findings emphasize the adverse impact of high microbial translocation on gut immune recovery and the necessity of establishing a novel therapeutic intervention to inhibit microbial translocation.

Thymic Function during 12 Months of Highly Active Antiretroviral Therapy in Thai HIV-Infected Patients with Normal and Slow Immune Recovery.

The aim of this study was to determine and compare thymic output during 12 months of highly active antiretroviral therapy (HAART) in HIV-infected patients with different types of immune recovery. In total, 18 Thai HIV-infected patients with normal immune recovery (NR) and 13 Thai HIV-infected patients with slow immune recovery (SR) were enrolled. T-cell receptor rearrangement excision circle (TREC) levels in peripheral blood mononuclear cells (PBMCs) and CD4(+) T cells were quantified at baseline, and after 6 and 12 months of HAART. CD4(+) T-cell counts in NR patients were significantly higher than those in SR patients after 6 and 12 months of HAART. However, the median TREC levels in PBMCs and CD4(+) T cells in both groups were comparable. Moreover, TREC levels showed similar trends in PBMCs and CD4(+) T cells in both groups during 12 months of HAART. Only patients with SR had significant increases in median TREC levels in PBMCs and CD4(+) T-cells during the first 6 months of HAART. No correlations were found between CD4(+) T-cell count and TREC levels in PBMCs and CD4(+) T cells. These results imply that the increase in CD4(+) T-cell count in SR patients after 12 months of HAART is likely attributable to thymic output and other sources.

HAART has no major impact on hematological and plasma bilirubin changes in HIV-infected patients with congenital G-6-PD deficiency.

Hematological effects of antiretroviral (ARV) drugs in HIV-infected patients with glucose-6-phosphate dehydrogenase (G-6-PD) deficiency are unclear. The aim of this study was to assess effects of highly active antiretroviral therapy (HAART) on hematological and plasma bilirubin changes in these patients. A hundred and nine HIV-infected Thai patients were tested for G-6-PD deficiency and its variant by using fluorescent spot test and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, accordingly. Changes of hematological parameters and plasma bilirubin at baseline and 6 months of HAART were analyzed in G-6-PD deficiency patients. G-6-PD deficiency was found in 10 (9.17%) patients and G-6-PD Canton1376G>T was the most frequent. Of these, 3 patients had coinheritance of G-6-PD deficiency and thalassemia. Increased mean levels of lymphocyte counts, CD4⁺ T-cells, mean corpuscular hemoglobin (MCH) and hemoglobin from baseline to 6 months of HAART were observed. Whereas, mean levels of total bilirubin and direct bilirubin at baseline were not significantly different from those at 6 months of HAART. Therefore, HAART did not cause hemolytic anemia and hyperbilirubinemia in HIV-infected patients with G-6-PD deficiency. On the other hand, the effective use of HAART is associated with improvements in hemoglobin and MCH levels of these patients.

Diagnosis of alpha-thalassemia-1 Southeast Asian type deletion and fetal gender by single-tube multiplex real-time PCR.

BACKGROUND: The combination of an x-linked hematologic disorder with a heterozygous alpha-thalassemia-1 Southeast Asian (SEA) type deletion might lead to severe anemia in male infants. This study is to develop a simple and cost-effective single tube multiplex real-time PCR for the diagnosis of alpha-thalassemia-1 SEA type deletion and detect fetal gender. METHODS: Multiplex real-time polymerase chain reaction (PCR) for the detection of alpha-thalassemia-1 SEA type deletion gene, wild type alpha-globin gene, and sex-determining region Y (SRY) gene was validated by analysis of 95 cord blood samples (60 normal individuals, 28 heterozygous of alpha-thalassemia-1 SEA type deletion and 7 Bart's hydrops fetalis). The change in threshold cycle (deltaC(T)) was analyzed by subtracting the C(T-mutant) from C(T-wild) type. RESULTS: Mean deltaC(T) values were significantly different among these three groups, and a SRY gene determination was 100% in concordance with fetal genders. Furthermore, analysis of fetal gender did not affect the deltaC(T) of alpha-thalassemia-1 SEA detection. CONCLUSIONS: Combined alpha-thalassemia-1 SEA type detection and fetal gender determination in a single-tube multiplex real-time PCR is an alternative assay for a conventional method for the diagnosis of alpha-thalassemia-1 SEA deletion and fetal gender.

Detection of α-thalassemia-1 Southeast Asian and Thai type deletions and ß-thalassemia 3.5-kb deletion by single-tube multiplex real-time PCR with SYBR Green1 and high-resolution melting analysis.

BACKGROUND: Prevention and control of thalassemia requires simple, rapid, and accurate screening tests for carrier couples who are at risk of conceiving fetuses with severe thalassemia. METHODS: Single-tube multiplex real-time PCR with SYBR Green1 and high-resolution melting (HRM) analysis were used for the identification of α-thalassemia-1 Southeast Asian (SEA) and Thai type deletions and ß-thalassemia 3.5-kb gene deletion. The results were compared with those obtained using conventional gap-PCR. DNA samples were derived from 28 normal individuals, 11 individuals with α-thalassemia-1 SEA type deletion, 2 with α-thalassemia-1 Thai type deletion, and 2 with heterozygous ß-thalassemia 3.5-kb gene deletion. RESULTS: HRM analysis indicated that the amplified fragments from α-thalassemia-1 SEA type deletion, α-thalassemia-1 Thai type deletion, ß-thalassemia 3.5-kb gene deletion, and the wild-type ß-globin gene had specific peak heights at mean melting temperature (T(m)) values of 86.89℃, 85.66℃, 77.24℃, and 74.92℃, respectively. The results obtained using single-tube multiplex real-time PCR with SYBR Green1 and HRM analysis showed 100% consistency with those obtained using conventional gap-PCR. CONCLUSIONS: Single-tube multiplex real-time PCR with SYBR Green1 and HRM analysis is a potential alternative for routine clinical screening of the common types of α- and ß-thalassemia large gene deletions, since it is simple, cost-effective, and highly accurate.