Global assessment of dengue Virus-Specific CD4+ T cell responses in Dengue-Endemic areas

Background: Dengue is a major public health problem worldwide. Assessment of adaptive immunity is important to understanding immunopathology and to define correlates of protection against dengue virus (DENV). To enable global assessment of CD4+ T cell responses, we mapped HLA-DRB1-restricted DENV-specific CD4+ T cell epitopes in individuals previously exposed to DENV in the general population of the dengue-endemic region of Managua, Nicaragua. Methods: HLA class II epitopes in the population of Managua were identified by an in vitro IFNγ ELISPOT assay. CD4+ T cells purified by magnetic bead negative selection were stimulated with HLA-matched epitope pools in the presence of autologous antigen-presenting cells, followed by pool deconvolution to identify specific epitopes. The epitopes identified in this study were combined with those previously identified in the DENV endemic region of Sri Lanka, to generate a “megapool” (MP) consisting of 180 peptides specifically designed to achieve balanced HLA and DENV serotype coverage. The DENV CD4MP180 was validated by intracellular cytokine staining assays. Results: We detected responses directed against a total of 431 epitopes, representing all 4 DENV serotypes, restricted by 15 different HLA-DRB1 alleles. The responses were associated with a similar pattern of protein immunodominance, overall higher magnitude of responses, as compared to what was observed previously in the Sri Lanka region. Based on these epitope mapping studies, we designed a DENV CD4 MP180 with higher and more consistent coverage, which allowed the detection of CD4+ T cell DENV responses ex vivo in various cohorts of DENV exposed donors worldwide, including donors from Nicaragua, Brazil, Singapore, Sri Lanka, and U.S. domestic flavivirus-naïve subjects immunized with Tetravalent Dengue Live-Attenuated Vaccine (TV005). This broad reactivity reflects that the 21 HLA-DRB1 alleles analyzed in this and previous studies account for more than 80% of alleles present with a phenotypic frequency ≥5% worldwide, corresponding to 92% phenotypic coverage of the general population (i.e., 92% of individuals express at least one of these alleles). Conclusion: The DENV CD4 MP180 can be utilized to measure ex vivo responses to DENV irrespective of geographical location

Fibrosing Blepharo-conjunctivitis following Pyogenic Granuloma in Ocular Acne Rosacea

Purpose: To report a case of pyogenic granuloma associated with ocular acne rosacea. Design: Case report. Methods: Interventional case report. Results: The patient developed an unusual lesion of the conjunctiva and limbus, histologically confirmed to be pyogenic granuloma. Excision of the lesion resulted in symblepharon formation, adhesions between lid margins, tarsal conjunctiva, and cornea, and reductions in visual acuity. The patient underwent surgical separation of the corneal lid adhesions with a mucosal autograft for the tarsal conjunctiva and an amniotic membrane graft for the cornea with good results. Conclusions: Ocular rosacea is a common condition but such excessive and unusual fibrotic healing responses following excision of pyogenic granulomas have not been previously reported. This case demonstrated significant morbidity and decreased visual acuity. However, early recognition and effective surgical management can lead to good visual outcomes

Evaluation of global differential gene and protein expression in primary pterygium: S100A8 and S100A9 as possible drivers of a signaling network

10.1371/journal.pone.0097402PLoS ONE95e9740

Cloning and pharmacological characterization of the dog histamine H4 receptor

The histamine