Summary B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin www.biorxiv.org
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One Line
B cells and HSV-specific antibodies play a significant role in controlling HSV-2 reactivation in the skin, with memory B cells and plasmablasts infiltrating genital skin during the tissue-based immune response, and the concentration of HSV-2-specific antibody in tissue varies independently from the concentration in serum.
Key Points
- B cells and antibody-secreting cells are present in skin biopsies of persons with symptomatic HSV-2 reactivation, with infiltrates at the greatest density at the time of reactivation.
- CD20+ and IgG RNA+ cells tend to cluster in genital skin during HSV-2 reactivation.
- The concentration of HSV-2-specific antibody in tissue varies independently from the concentration in serum, suggesting local mechanisms.
- HSV-specific antibodies are present in the skin during reactivation, and the peak of cell density is shown by persons with healing time.
- The study provides a new understanding of the immune response to HSV-2 reactivation in the skin and highlights the importance of B cells and HSV-specific antibodies in controlling the virus.
Summaries
342 word summary
B cells and HSV-specific antibodies respond to herpes simplex virus type 2 (HSV-2) reactivation in the skin, with memory B cells and plasmablasts infiltrating genital skin during the tissue-based immune response. Antibody-secreting cells were found clustered in an area of dense T-cell infiltration, with CD20+ cells per mm2 correlating with CD8+ cells per mm2 and CD4+ cells per mm2. The concentration of HSV-2-specific antibody in tissue varies independently from the concentration in serum, suggesting local mechanisms. The study provides a new understanding of the immune response to HSV-2 reactivation in the skin and highlights the importance of B cells and HSV-specific antibodies in controlling the virus. A study analyzed B cells and antibodies in skin extracts from 14 participants during HSV-2 reactivation. The concentration of HSV-2-specific IgG varied over time and by individual, with some participants having a peak at the lesion and others during early healing. Two patterns of B-cell infiltration are seen, with different subsets of B cells possibly regulating inflammation. IgG against gB2 and gD2 is present at higher levels than non-HSV antigens. The study investigated the presence and distribution of antibody-secreting cells (ASCs) in response to HSV-2 reactivation and skin healing. The majority of observed CD20+ cells were not memory B cells and that T cells are often observed to be present near hair follicles or skin-localized glandular structures with little to no overlap interactions between the two subpopulations of cells. Biopsies of active HSV-2 showed the greatest extent of B cells and IgG in the upper dermis of lesion and early healing tissue biopsy sections. CD4+ and CD20+ cells tend to cluster in genital skin during HSV-2 reactivation. The density of CD20+ and IgG RNA+ cells was considerably lower than T-cell density at the site of healing. The peak in antibody titer was at 8 weeks post-healing time point. All 16 participants were seropositive for HSV-2 and 6 also had antibody to HSV-1. The investigation into the role of B cells and ASCs in local inflammatory processes confirms the importance of spatially-informative methods and urges further exploration.
878 word summary
A study conducted by Johnston, Wald, Zhu, and Corey examines the role of B cells and HSV-specific antibodies in response to HSV-2 reactivation in non-mucosal skin. The study demonstrates that B cells and antibody-secreting cells are present in skin biopsies of persons with symptomatic HSV-2 reactivation, with infiltrates at the greatest density at the time of reactivation. The presence and role of B cells and ASCs in the immune response to HSV-2 reactivation in non-mucosal skin is explored using genital skin biopsies. B cells and ASCs have been confirmed to be present in skin through single-cell and RNAseq technology, as well as in inflammatory infiltrates related to HSV. B cells were identified by CD20 immunofluorescence in the upper dermis and were distributed within areas of immune cell infiltration. They were most often observed to be located in dense groups of T cells. All 16 participants were seropositive for HSV-2 and 6 also had antibody to HSV-1. The investigation into the role of B cells and ASCs in local inflammatory processes confirms the importance of spatially-informative methods and urges further exploration. A study investigated the presence and distribution of antibody-secreting cells (ASCs) in response to HSV-2 reactivation and skin healing. The study found that the majority of observed CD20+ cells were not memory B cells and that T cells are often observed to be present near hair follicles or skin-localized glandular structures with little to no overlap interactions between the two subpopulations of cells. Biopsies of active HSV-2 showed the greatest extent of B cells and IgG in the upper dermis of lesion and early healing tissue biopsy sections. CD4+ and CD20+ cells tend to cluster in genital skin during HSV-2 reactivation. The density of CD20+ and IgG RNA+ cells was considerably lower than T-cell density at the site of healing. The peak in antibody titer was at 8 weeks post-healing time point. A study analyzed B cells and antibodies in skin extracts from 14 participants during HSV-2 reactivation. The concentration of HSV-2-specific IgG varied over time and by individual, with some participants having a peak at the lesion and others during early healing. The study found that the immunoglobulin observed in biopsies was specific to HSV-2, and that the IgG detected in skin during HSV-2 reactivation was produced by an antigen-specific process. Two patterns of B-cell infiltration are seen, with different subsets of B cells possibly regulating inflammation. IgG against gB2 and gD2 is present at higher levels than non-HSV antigens. The role of B cells or ASCs in this process is largely unknown, but there is a boosting of local antibody secretion due to viral challenge where tissue-based antibody concentration is increased during HSV-2-reactivation and healing, and much lower in uninvolved control tissues. Confirmation by CD27 co-staining that some of these B cells are of a memory phenotype is therefore not reliable by IF, and observation of co-expression by FISH was necessary. Mechanistic evaluation from human tissues is by necessity limited, and as such this study was limited in scope to those persons available and willing to undergo serial biopsies and blood draws over the course of symptomatic HSV-2 infection. A study on healthy adults with a history of symptomatic genital herpes found B cells and antibody-secreting cells present in the inflammatory infiltrate resulting from HSV-2 reactivation in genital-region skin. The concentration of HSV-2-specific antibody in tissue varies independently from the concentration in serum, suggesting local mechanisms. The study used biopsy extracts to study B cells and HSV-specific antibodies in response to HSV-2 reactivation in skin, with increased antibody concentration and cell density observed. The article concludes that the study provides a new understanding of the immune response to HSV-2 reactivation in the skin and highlights the importance of B cells and HSV-specific antibodies in controlling the virus. B cells and HSV-specific antibodies respond to HSV-2 reactivation in the skin. The study found that B cells and antibodies respond to herpes simplex virus type 2 (HSV-2) reactivation in the skin. B cells, including memory B cells and plasmablasts, infiltrate genital skin during the tissue-based immune response to HSV-2 reactivation. HSV-specific antibodies are present in the skin during reactivation. Multiple biopsies were taken from the genital area of symptomatic participants, and CD20+ and IgG RNA+ cells were measured from biopsies over time. Antibody-secreting cells were found clustered in an area of dense T-cell infiltration. The peak of cell density was shown by persons with healing time and shown by their IF in tissue over time. CD20+ cells per mm2 were found to have a correlation with CD8+ cells per mm2 and CD4+ cells per mm2. A study was conducted to examine the response of B cells and HSV-specific antibodies to HSV-2 reactivation in skin. The study included participant demographics and HSV clinical history, and used immunofluorescent detection of IgG in genital skin biopsies in one subject. The study found that B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin, and that the levels of HSV-2 specific antibodies did not change during HSV-2 reactivation. Serum samples were collected from participants at different time points, and specific IgG levels were measured for HSV-2, Epstein-Barr virus (EBV) gp350, and influenza hemagglutinin (Flu-HA). The study also included area under the curve (AUC) measurements of serum samples collected at participant enrollment.
2589 word summary
The document reports on a study of B cells and herpes simplex virus (HSV)-specific antibodies in response to HSV-2 reactivation in skin. Serum samples were collected from participants at different time points, and specific IgG levels were measured for HSV-2, Epstein-Barr virus (EBV) gp350, and influenza hemagglutinin (Flu-HA). The study found that the levels of HSV-2 specific antibodies did not change during HSV-2 reactivation. The study also included area under the curve (AUC) measurements of serum samples collected at participant enrollment. Serum dilution reciprocal was measured for each participant at different time points. The study found that B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin. The study examines the response of B cells and HSV-specific antibodies to HSV-2 reactivation in skin. The levels of HSV-2 specific antibody and total IgG follow B-cell and ASC density, and IgG levels are based on the immune response to HSV-2 reactivation. Antibody is detectable during the tissue-based immune response to HSV-2 reactivation, and the amount is variable over time. The study also includes participant demographics and HSV clinical history. The sample series is divided by time of peak over time, and all lesion site biopsies are included in each graph. The study uses immunofluorescent detection of IgG in genital skin biopsies in one subject, and CD4+, CD8+, and CD20+ cells are included in each graph. Missing data points correspond to biopsies with insufficient tissue to run the test. B cells and HSV-specific antibodies respond to HSV-2 reactivation in the skin. Multiple biopsies were taken from the genital area of symptomatic participants. CD20+ and IgG RNA+ cells were measured from biopsies over time, with counts taken from the healing lesion and the control lesion. There was no correlation between B and T cell subsets or between B cell and T cell counts. The peak of cell density was shown by persons with healing time and shown by their IF in tissue over time. IgG+ cells were found infiltrating tissue without relation to small capillaries, adjacent to small capillaries, and within small capillaries. Antibody-secreting cells were found clustered in an area of dense T-cell infiltration. CD20+ cells per mm2 were found to have a correlation with CD8+ cells per mm2 and CD4+ cells per mm2. B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin. Cells expressing high levels of IgG RNA infiltrate genital skin during the tissue-based immune response to HSV-2 reactivation. B cells infiltrate genital skin during the tissue-based immune response to HSV-2 reactivation, as shown by CD20 + CD79b + B cells in serial genital skin biopsies. T cells are also present in intra-dermal glandular structures and dense clusters, as well as diffuse intradermal infiltration. Real-time taqman PCR assay can detect HSV DNA on mucosal surfaces. RNAscope is a novel in situ RNA analysis platform for formalin-fixed, paraffin-embedded tissues. Image analysis can accurately count CD4+ and CD8+ T cells in human tissue. B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin. The study examined the response of B cells and antibodies to herpes simplex virus type 2 (HSV-2) reactivation in the skin. The research found that B cells, including memory B cells and plasmablasts, responded to the reactivation of HSV-2 in the skin. The study also found that HSV-specific antibodies were present in the skin during reactivation. These findings suggest that the skin is a novel niche for recirculating B cells, which may play a role in the immune response to HSV-2 reactivation in the skin. B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin. Studies have shown that B cells play a role in skin simplex lesions and that IgM, IgG1, IgG3, and IgA reach the human fibroblast populations in human skin. Resident memory CD8 T cells trigger protective innate and adaptive immune responses. Maternal antiviral immunoglobulin accumulates in neural tissue of neonates to protect male mice from lethal skin challenge with clinical isolates of HSV-1 and HSV-2. An HSV-2 single-cycle candidate vaccine deleted in glycoprotein D, ?gD-2, protects against HSV-2 infection in HSV-2 seronegative women. Therapeutic vaccine GEN-003 results in durable reduction in genital lesions. Immune responses elicited by GEN-003 candidate HSV-2 therapeutic vaccine in a randomized controlled dose-ranging phase 1/2a trial. Dissection of the antibody response against herpes simplex virus glycoproteins in naturally infected humans. A local macrophage chemokine network sustains protective tissue-resident memory CD4 T cells. This article discusses the response of B cells and HSV-specific antibodies to HSV-2 reactivation in the skin. Several studies have shown that herpes simplex virus (HSV) can cause genital tract shedding episodes and poor immunologic control in ganglia and genital mucosa. The study design involved participant recruitment, clinical guidance, tissue biopsy analysis, and data analysis. The results showed that B cells and ASCs were present in the skin during HSV-2 reactivation, and their density was highest at the maximum of lesion or newly healed. Serum antibody titer over time was also compared, and statistical significance was defined as adjusted, two-sided p value ?0.05. The article concludes that the study provides a new understanding of the immune response to HSV-2 reactivation in the skin and highlights the importance of B cells and HSV-specific antibodies in controlling the virus. This study examines the response of B cells and HSV-specific antibodies to HSV-2 reactivation in skin. The study used quantitative PCR to measure HSV-2 DNA in biopsy specimens and genital swabs, and Luminex binding antibody assay to measure HSV-2 specific antibodies. The data was analyzed using R Studio and GraphPad Prism. The level of background was assigned using serum samples from ten HSV-2 seropositive and ten HSV-2 seronegative donors. The study found that B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin, with increased antibody concentration and cell density observed. The document discusses the use of biopsy extracts to study B cells and HSV-specific antibodies in response to HSV-2 reactivation in skin. The extracts were prepared by placing freshly sliced biopsy cryosections in a microcentrifuge tube with Tissue Extraction Reagent II and extracting protein at room temperature. Fluorescent in situ hybridization (FISH) was used to identify the presence of IgG-producing cells in biopsies and determine their spatial localization and interaction with other cell types. Antibodies were applied after non-specific binding was blocked, and the relative density of B cells and/or antibody in tissue biopsy sections was determined using a cell counter plugin. The study was not certified by peer review and is available under a CC-BY-NC-ND 4.0 International license. A study conducted on healthy, HSV-2-seropositive adults with a history of symptomatic genital herpes showed the presence of B cells and antibody-secreting cells (ASCs) in the inflammatory infiltrate resulting from HSV-2 reactivation in genital-region skin. The kinetics of tissue infiltration and egress of CD20+ cells, but less so IgG+ cells, follows that seen in T cell studies. The concentration of HSV-2-specific antibody in tissue varies independently from the concentration in serum, suggesting that there are local mechanisms either leading to local production or recruitment of HSV-2-specific antibody. Further investigation is needed to identify markers of tissue infiltration and recruitment of B cells, which has been proposed to be necessary for the immune response to HSV-2 reactivation. B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin. Evidence from FISH and IF suggest that multiple B-cell types may be involved in the inflammatory or healing process after symptomatic HSV-2 reactivation. The role of B cells or ASCs in this process is largely unknown, but there is a boosting of local, if not systemic, antibody secretion due to viral challenge where tissue-based antibody concentration is increased during HSV-2-reactivation and healing, and much lower in uninvolved control tissues. The finding is congruous with observations in other conditions where a peak in ASC density is offset in time to the peak in corresponding antibody production. Confirmation by CD27 co-staining that some of these B cells are of a memory phenotype is therefore not reliable by IF, and observation of co-expression by FISH was necessary. It is possible that tissue-infiltrating memory cells may give rise to ASCs during the course of HSV-2 reactivation and healing; this needs to be tested by B-cell receptor sequencing. Mechanistic evaluation from human tissues is by necessity limited, and as such this study was limited in scope to those persons available and willing to undergo serial biopsies and blood draws over the course of symptomatic HSV-2 infection. B cells and antibodies specific to HSV-2 are present in skin biopsies during reactivation. Two patterns of B-cell infiltration are seen, with different subsets of B cells possibly regulating inflammation. The presence of CD20+ B cells in T-cell clusters suggests early stage B cells involved in antigen processing. The study shows multiple mechanisms of antibody infiltration into inflamed skin, including local production and antigen-dependent transudation from serum. The dynamics of HSV-specific antibody in skin and serum differ from control and EBV-specific antibodies. IgG against gB2 and gD2 is present at higher levels than non-HSV antigens. The study suggests both tissue antibody production and antigen-driven B-cell recruitment mechanisms contribute to the discrepancy between HSV-specific and control antigens in skin and serum. The study analyzed B cells and antibodies in skin extracts from 14 participants during HSV-2 reactivation. The concentration of HSV-2-specific IgG varied over time and by individual, with some participants having a peak at the lesion and others during early healing. The difference in the amount of HSV-specific antibody also varied by up to 30-fold within participants. The study found that the immunoglobulin observed in biopsies was specific to HSV-2, and that the IgG detected in skin during HSV-2 reactivation was produced by an antigen-specific process. The systemic antibody response during HSV-2 reactivation was consistent with earlier reports documenting the absence of a systemic antibody response. The concentration of IgG detected in skin during HSV-2 reactivation is produced by locally-migrated B cells and is therefore present at higher concentration than non-HSV-specific antibody. A study found that B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin. Biopsies of active HSV-2 showed the greatest extent of B cells and IgG in the upper dermis of lesion and early healing tissue biopsy sections. The lymphocyte subsets may be influenced by similar recruitment mechanisms. CD4+ and CD20+ cells tend to cluster in genital skin during HSV-2 reactivation. The density of CD20+ and IgG RNA+ cells was considerably lower than T-cell density at the site of healing. In three persons, few B cells were seen in any biopsies. The CD20+ cell density of ulcerative or healing lesions was higher than the maximum of the 2, 4, or 8 weeks post-healing biopsies. Similarly, the peak in antibody titer was at 8 weeks post-healing time point. A study found that B cells and HSV-specific antibodies respond to HSV-2 reactivation in genital skin. The density of CD20+ B cells peaked at the time of healing, about 10 days after lesion onset, with a median of 18.4 cells/mm2. The highest density of CD20+ B cells was observed in biopsies when genital lesions were present. The magnitude and density of the B-cell and ASC infiltrate were greatest during the acute phases of reactivation, during clinical lesion and early healing. Dermal-infiltrating and capillary-associated B cells and ASCs may all be related to HSV-2 reactivation in genital skin. Clusters of T cells were also observed in the dermis containing both CD8+ and CD4+ T cells. The observed B cells and ASCs may have migrated into the tissue due to localization in small capillaries. CD138 and IgG signal were concomitantly present in single cells, confirming that these cells are antibody-secreting. The study investigated the presence and distribution of antibody-secreting cells (ASCs) in response to HSV-2 reactivation and skin healing. Fluorescent in situ hybridization (FISH) directed at IgG mRNA was performed in 57 biopsies from 10 individuals to identify cells with dense expression of immunoglobulin transcripts. Punctate fluorescence indicative of ASCs was observed in 48 of the 57 tested biopsies. CD27, a canonical marker of memory B cells, is occasionally observed in staining of B cells, but CD20 + cells are not memory B cells. The overall frequency of PAX5 detection was low, and PAX5 nuclear signal was also seen in cells not containing IgG transcripts. The co-localization of a high level of PAX5 with a low level of IgG transcript can indicate memory B and memory lineage B cells but is downregulated by 4-5 fold in ASCs. Nuclear IRF4 production was frequently observed to be co-localized with IgG transcripts by confocal microscopy. The study found that the majority of observed CD20 + cells were not memory B cells and that T cells are often observed to be present near hair follicles or skin-localized glandular structures with little to no overlap interactions between the two subpopulations of cells. B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin. B cells were identified by CD20 immunofluorescence in the upper dermis and were distributed within areas of immune cell infiltration. They were most often observed to be located in dense groups of T cells. B cells were present in all of the biopsies from the HSV-2 lesion site and were detected during the lesion and early healing, similarly to T-cell infiltration. The number of CD20/CD79b+ cells present in each biopsy section varied by person and timepoint. In general, B cells were more common than CD19. Dual staining was employed to assist in identification of non-specific antibody binding. The most common site of biopsies was the buttock. All 16 participants were seropositive for HSV-2 and 6 also had antibody to HSV-1. The investigation into the role of B cells and ASCs in local inflammatory processes confirms the importance of spatially-informative methods and urges further exploration. The presence and role of B cells and ASCs in the immune response to HSV-2 reactivation in non-mucosal skin is explored using genital skin biopsies. B cells and ASCs have been confirmed to be present in skin through single-cell and RNAseq technology, as well as in inflammatory infiltrates related to HSV. Animal models indicate that B cells are important for protection from viral infections, including HSV-2, and B cell-deficient mice rapidly develop severe disease after HSV-2 genital challenge. The role of humoral immunity in the human response to HSV-2 reactivation is less clear, as infiltrating or resident memory CD4 + T cells are important factors in local viral control. Certain HSV-specific antibodies may function protectively, potentially due to lymphocyte recruitment by VCAM-1. The study examines the role of B cells and HSV-specific antibodies in response to HSV-2 reactivation in skin. While cell-mediated immunity is important, the host's adaptive immune response is thought to be the primary mechanism for controlling viral reactivation. The spectrum of clinical disease among infected persons is broad, with some experiencing asymptomatic seroconversion and others having 10 or more recurrences a year. Tissue-based T cells have been shown to be important effectors in controlling mucosal viral infections, while tissue-based B cells' role is less known. The study demonstrates that B cells and antibody-secreting cells are present in skin biopsies of persons with symptomatic HSV-2 reactivation, with infiltrates at the greatest density at the time of reactivation. HSV-2-specific antibodies to surface antigens are also detected. The study suggests a functional and distinct role of tissue-based B cells in the local immune response to HSV-2. B cells and HSV-specific antibodies respond to HSV-2 reactivation in skin. The study was conducted by a team of authors, including Johnston, Wald, Zhu, and Corey. The preprint is available under a CC-BY-NC-ND 4.0 International license and was posted on July 8, 2020. The study was not certified by peer review.