Over the last two decades, there have been three deadly human outbreaks of coronaviruses (CoVs) caused by SARS-CoV, MERS-CoV, and SARS-CoV-2, which has caused the current COVID-19 global pandemic. All three deadly CoVs originated from bats and transmitted to humans via various intermediate animal reservoirs. It remains highly possible that other global COVID pandemics will emerge in the coming years caused by yet another spillover of a bat-derived SARS-like coronavirus (SL-CoV) into humans. Determining the Ag and the human B cells, CD4 1 and CD8 1 T cell epitope landscapes that are conserved among human and animal coronaviruses should inform in the development of future pan-coronavirus vaccines. In the current study, using several immunoinformatics and sequence alignment approaches, we identified several human B cell and CD4 1 and CD8 1 T cell epitopes that are highly conserved in 1) greater than 81,000 SARS-CoV-2 genome sequences identified in 190 countries on six continents; 2) six circulating CoVs that caused previous human outbreaks of the common cold; 3) nine SL-CoVs isolated from bats; 4) nine SL-CoV isolated from pangolins; 5) three SL-CoVs isolated from civet cats; and 6) four MERS strains isolated from camels. Furthermore, the identified epitopes: 1) recalled B cells and CD4 1 and CD8 1 T cells from both COVID-19 patients and healthy individuals who were never exposed to SARS-CoV-2, and 2) induced strong B cell and T cell responses in humanized HLA-DR1/HLA-A*02:01 double-transgenic mice. The findings pave the way to develop a preemptive multiepitope pancoronavirus vaccine to protect against past, current, and future outbreaks.

Hepatitis E virus (HEV) causes acute viral hepatitis and is endemic in the developing world. Few data are available on cellular immune responses in HEV infection. Using flow cytometry, we studied the frequencies of peripheral blood CD4 + /CD8 + T cells secreting interferon (IFN)-γ, tumor necrosis factor (TNF)-α, and interleukin (IL)-4 in 21 patients with acute hepatitis E and 18 healthy controls, after stimulation with the HEV capsid (ORF2) protein. Cytokine levels in serum specimens and culture supernatants of ORF2-stimulated peripheral blood mononuclear cells (PBMCs) were estimated in enzyme-linked immunosorbent assays. In addition, cytokine mRNA transcripts were measured in PBMCs by reverse transcription-polymerase chain reaction. In patients with acute hepatitis E, although the total CD4 + population was expanded, the proportions of CD4 + /CD69 + and CD8 + /CD69 + cells producing IFN-γ, TNF-α, and IL-4 in response to HEV ORF2 stimulation were unchanged. However, IFN-γ levels in the supernatants and IFN-γ mRNA transcripts in cells were elevated in ORF2-stimulated PBMCs in acute hepatitis E; levels of IL-2 or TNF-α were unchanged. Our findings suggest that CD4 + IFN-γ-secreting cells, which do not belong either to the helper T cell type 1 or type 2 phenotype, as is the case with natural killer T cells, may be involved in the pathogenesis of hepatitis E. Further, the limited immune reactivity we detected in peripheral blood cells may be related to the sequestration of immune events to the intrahepatic compartment, which is the major disease site.

Objective:To evaluate the efficacy of coronally advanced flap (CAF) procedure under microsurgical approach for the management of Miller's Class I and II gingival recession defects with the use of either platelet-rich fibrin (PRF) or amnion membrane (AM) in comparison to CAF alone.Materials and Methods:A total of 45 sites with Miller's Class I or II gingival recession defect were randomly distributed for: Experimental Group I (CAF with PRF) sites (n = 15) which were treated with the microsurgical approach using CAF along with PRF; experimental Group II (CAF with AM) sites (n = 15) were treated with the microsurgical approach using CAF along with AM; control Group III (CAF alone) sites (n = 15) were treated with the microsurgical approach using CAF alone. Vertical gingival recession (VGR), horizontal gingival recession (HGR), gingival thickness (GT) (using transgingival probing [TGP] and ultrasonography [USG]) and patients’ response and acceptance were documented at baseline, 3 months and 6 months after surgical interventions.Results:CAF alone and in combination with PRF or AM, were effective techniques for root coverage with average VGR values of 1.47 ± 0.92 mm (56%), 0.67 ± 1.23 mm (36%) and 0.60 ± 1.06 mm (33%) in Group I (CAF with PRF), Group II (CAF with AM), and Group III (CAF alone), respectively. Complete coverage (100%) was obtained in 33.3% sites of Group I (CAF with PRF), 26.6% sites of Group II (CAF with AM) and 13.3% in Group III (CAF alone). Patients’ response and acceptance for surgical treatment modality in terms of patient esthetic score and decrease in hypersensitivity score was highest for Group I (CAF with PRF), whereas patient comfort score was highest for Group II (CAF with AM). At 6 months follow-up, significant increase in GT measurements (using TGP and USG) in Group I (CAF with PRF), whereas, nonsignificant increase for Group II (CAF with AM) and no change or decrease for Group III (CAF alone) as compared to baseline was observed.Conclusion:The present study observed enhancement in root coverage when PRF or AM are used in conjunction with CAF as compared to CAF alone. These results are based on 6-month follow-up. Therefore, the long-term evaluation may be necessary to appreciate the clinical effect of autologous PRF and AM.

The Herpes Simplex Virus type 1 virion tegument phosphoprotein 11/12 (HSV-1 VP11/12) is a major antigen targeted by CD8+ T cells from HSV-seropositive individuals. However, whether and which VP11/12-epitope-specific CD8+ T cells play a role in the “natural” protection seen in seropositive healthy asymptomatic (ASYMP) individuals (who have never had clinical herpes disease) remain to be determined. In this study, we used multiple prediction computer-assisted algorithms to identify 10 potential HLA-A*02:01-restricted CD8+ T cell epitopes from the 716 amino acids sequence of VP11/12. Three out of ten epitopes exhibited high to moderate binding affinity to HLA-A*02:01 molecules. In ten sequentially studied HLA-A*02:01 positive and HSV-1-seropositive ASYMP individuals, the most frequent, robust and polyfunctional effector CD8+ T-cell responses, as assessed by a combination of tetramer frequency, granzyme B, granzyme K, perforin, CD107a/b cytotoxic degranulation, IFN-γ and multiplex cytokines assays, were predominantly directed against three epitopes: VP11/1266–74, VP11/12220–228 and VP11/12702–710. Interestingly, ASYMP individuals had significantly higher proportion of CD45RAlowCCR7lowCD44highCD62LlowCD27lowCD28lowCD8+ effector memory T cells (TEM) specific to the three epitopes, compared to symptomatic (SYMP) individuals (with a history of numerous episodes of recurrent ocular herpetic disease). Moreover, immunization of HLA-A*02:01 transgenic mice with the three ASYMP CD8+ TEM cell epitopes induced robust and polyfunctional epitope-specific CD8+ TEM cells that were associated with a strong protective immunity against ocular herpes infection and disease. Our findings outline phenotypic and functional features of protective HSV-specific CD8+ T cells that should guide the development of an effective T-cell-based herpes vaccine.

Circulating conventional memory CD8 T cells (i.e., the CD8 effector memory T [T] cell and CD8 central memory T [T] cell subsets) and the noncirculating CD8 tissue-resident memory T (T) cell subset play a critical role in mucosal immunity. Mucosal chemokines, including the recently discovered CXCL17, are also important in mucosal immunity because they are homeostatically expressed in mucosal tissues. However, whether the CXCL17 chemokine contributes to the mobilization of memory CD8 T cell subsets to access infected mucosal tissues remains to be elucidated. In this study, we report that after intravaginal HSV type 1 infection of B6 mice, we detected high expression levels of CXCL17 and increased numbers of CD44CD62LCD8 T and CD103CD8 T cells expressing CXCR8, the cognate receptor of CXCL17, in the vaginal mucosa (VM) of mice with reduced genital herpes infection and disease. In contrast to wild-type B6 mice, the CXCL17 mice developed 1) fewer CXCR8CD8 T and T cells associated with more virus replication in the VM and more latency established in dorsal root ganglia, and 2) reduced numbers and frequencies of functional CD8 T cells in the VM. These findings suggest that the CXCL17/CXCR8 chemokine pathway plays a crucial role in mucosal vaginal immunity by promoting the mobilization of functional protective CD8 T and CD8 T cells, within this site of acute and recurrent herpes infection.

Most blinding ocular herpetic disease is due to reactivation of herpes simplex virus 1 (HSV-1 IMPORTANCESeventy percent to 90% of adults harbor herpes simplex virus 1 (HSV-1), which establishes lifelong latency in sensory neurons of the trigeminal ganglia. This latent state sporadically switches to spontaneous reactivation, resulting in viral shedding in tears. Most blinding herpetic disease in humans is due to reactivation of HSV-1 from latency rather than to primary acute infection. To date, there is no licensed therapeutic vaccine that can effectively stop or reduce HSV-1 reactivation from latently infected sensory ganglia and the subsequent shedding in tears. In the present study, we demonstrated that topical ocular therapeutic vaccination of latently infected HLA transgenic rabbits with a lipopeptide vaccine that contains exclusively human "asymptomatic" CD8 ؉ T-cell epitopes successfully decreased spontaneous HSV-1 reactivation, as judged by a significant reduction in spontaneous shedding in tears. The findings should guide the clinical development of a safe and effective T-cell-based therapeutic herpes vaccine.

Herpes simplex virus type 1 (HSV-1) is a prevalent human pathogen that infects over 3.72 billion individuals worldwide and can cause potentially blinding recurrent corneal herpetic disease. HSV-1 establishes latency within sensory neurons of trigeminal ganglia (TG) and TG-resident CD8+ T cells play a critical role in preventing its reactivation. The repertoire, phenotype and function of protective CD8+ T cells are unknown. Bolstering the apparent feeble numbers of CD8+ T cells in TG remains a challenge for immunotherapeutic strategies. In this study, a comprehensive panel of 467 HLA-A*0201-restricted CD8+ T cell epitopes were predicted from the entire HSV-1 genome. CD8+ T cell responses to these genome-wide epitopes were compared in HSV-1 seropositive symptomatic (SYMP) individuals (with a history of numerous episodes of recurrent herpetic disease) vs. asymptomatic (ASYMP) individuals (who are infected but never experienced any recurrent herpetic disease). Frequent polyfunctional HSV-specific effector memory IFN-γ+CD107a/b+CD44highCD62LlowCD8+ TEM cells were detected in ASYMP individuals and were mainly directed against three “ASYMP” epitopes. In contrast, SYMP individuals have more mono-functional central memory CD44highCD62LhighCD8+ TCM cells. Furthermore, therapeutic immunization with an innovative prime/pull vaccine, based on priming with multiple “ASYMP” epitopes (prime) and neurotropic TG delivery of the T-cell attracting chemokine CXCL-10 (pull), boosted the number and function of CD44highCD62LlowCD8+ TEM and tissue-resident CD103highCD8+ TRM cells in TG of latently infected HLA-A*0201 Tg mice and reduced recurrent ocular herpes following UV-B induced reactivation. These findings have profound implications in the development of T-cell-based immunotherapeutic strategies to treat blinding recurrent herpes infection and disease.

A significantly larger repertoire of dysfunctional (exhausted) HSV-specific CD8(+)T cells were found in the TG of HLA transgenic rabbits latently infected with wild-type HSV-1 or with LAT-rescued mutant (i.e., LAT(+)TG) than in a more restricted repertoire of functional HSV-specific CD8(+)T cells in the TG of HLA transgenic rabbits latently infected with LAT-null mutant (i.e., LAT(-)TG). These findings suggest that the HSV-1 LAT locus interferes with the host cellular immune response by shaping a broader repertoire of exhausted HSV-specific CD8(+)T cells within the latency/reactivation TG site.