bluuebunny/crossref_metadata_2025_split huggingface dataset:
αE Integrin Compensates for β7 Deficiency in the Nasal Passages (NP) Following Nasal Immunization with Cholera Toxin (CT) (41.10)
Authors: Maslanka Tomasz , Riccardi Carol , Pascual David W. 
Published: January 2023

Abstract
Nasal immunization stimulates robust Ab responses in the head and neck lymph nodes (HNLN) and nasal-associated lymphoid tissue (NALT). Studies showed that B lymphocyte trafficking in the HNLN and NALT is both L-selectin- and β7-dependent. β7 integrin generally associates with αE or α4 integrins to mediate lymphocyte retention in the epithelium or the mucosa, respectively. To study the contribution of β7 integrin to upper respiratory immunity, β7−/− mice were nasally dosed with CT on days 0, 7, and 14. Mucosal IgA or serum IgG Ab titers were not different from B6 β7+/+ mice at any of the time points examined til day 42. Vaginal IgG responses were slightly enhanced in β7−/− mice, and absent in L-selectin−/− mice. However, the absence of β7 did alter the types of supporting B cells found in the NP. Increased expression of αE was observed on day 42 despite the absence of its co-receptor, β7. This increased αE expression was not observed in B6 β7+/+ mice. αE+ B cells continued to be found in the β7−/− small intestinal lamina propria. Minimal αE+ B cells were found in the HNLN. From these studies, although mucosal Ab responses remain intact in β7−/− mice, it does appear that different homing receptor-dependent mechanisms are employed.
This work is supported by NIH AI-55563.
Using Overlapping Synthetic Peptides from Tumor-Associated Protein Antigens for Breast Cancer Vaccine Development. (41.11)
Authors: Mirshahidi Saied , Whitney James B. , Kramer Victor G. , Anderson Karen S. , Ruprecht Ruth M. 
Published: January 2023

Abstract
An effective vaccine will need to elicit both CD4+ T helper and CD8+ cytotoxic T lymphocytes (CTL) responses in order to maintain effective CTL functions. Peptide vaccines tailored to certain genetic backgrounds are capable of eliciting MHC Class I-restricted (CD8+) and Class II-restricted (CD4+) responses. However, the need to identify specific T-cell epitopes in the context of MHC alleles hampers this approach. We have developed a novel strategy that bypasses the need for epitope mapping for an outbred population: vaccination with overlapping synthetic peptides (OSP), to generate strong antigen-specific cell-mediated immunity (CMI) in individual vaccine recipients with different MHC backgrounds. OSP contain all possible epitopes for the CD4+ as well as CD8+ T cells of a genetically diverse population. BALB/c mice were immunized with OSP derived from a tumor protein, which is overexpressed in 60% in breast cancer specimens. The vaccinated animals developed an in vitro activity of tumor specific- CTL, proliferative response, IFN-g production and CD107 expression. Immunized mice also were challenged with TS/A tumor cells (a murine mammary adenocarcinoma cell) to induce micrometastasis. Vaccination with OSP significantly increased the animal life span. The OSP vaccine strategy provides a novel opportunity to induce cellular immune responses against developing tumors in a genetically diverse population.
Generation and maintenance of antigen-specific CD8 T cells in the vaginal mucosa following systemic versus mucosal routes of immunization (41.6)
Authors: Onami Thandi M. , Suvas Pratima Krishna , Dech Heather M. , Sambira Fleurette 
Published: January 2023

Abstract
In this study we compared the efficacy of two different routes of viral infection on the generation, maintenance, and localization of antigen specific CD8T cells in lymphoid, non-lymphoid, and mucosal tissues during peak effector and memory phases of an immune response. We used an adoptive transfer system of naïve transgenic CD8 T cells, followed by intra-peritoneal (i.p) or intra-nasal (i.n) LCMV infection. Surprisingly, on day 8 the peak of the primary response, i.p. infection resulted in higher numbers of antigen-specific CD8T cells in the vaginal mucosa and ILN only, in addition to 2–3x more antigen specific CD8 T cells that co-expressed both IFNγ and TNFα, when compared to the i.n. route of infection. During memory, equivalent numbers of antigen specific CD8T cells were detected at all sites, irrespective of the route of infection. Following intravaginal re-challenge, both i.p. and i.n generated memory CD8 T cells showed robust secondary responses in the vaginal tract, however the i.n. memory CD8 T cell response was consistently greater. These data suggest that both systemic and mucosal routes of infection can generate a similar quantity of memory CD8 T cells in mucosal sites. However, the initial route of infection appears to influence the programming of the memory CD8T cells generated, and these early differences in priming may impact the quality of protective secondary responses.
Antibody-Mediated Protection against Genital Herpes Simplex Virus type 2 Disease in Mice by Fc gamma Receptor -Dependent and -Independent Mechanisms (41.8)
Authors: Milligan Gregg N , Chu C-F. , Meador M. G. , Young C. G. , Strasser J. E. , Bourne N. 
Published: January 2023

Abstract
The requirement for Ab -Fcγ receptor (FcγR) interactions or for virus neutralization in protection against genital HSV-2 challenge was examined. Serum IgG Ab isolated from HSV-immune mice protected normal mice against HSV-2 disease when administered prior to challenge. However, protection was significantly diminished in mice lacking the γ chain subunit utilized in FcγRI, FcγRIII, FcγRIV, and FcεRI and in normal mice depleted of FcγR+, Gr-1+ immune cells suggesting protection was largely mediated by an FcγR-dependent mechanism. To test if FcγR-independent antibody-mediated mechanisms might manifest protection differently, a highly neutralizing, HSV glycoprotein D -specific monoclonal antibody (mAb) was utilized. Administration of IgG1, IgG2a, or IgG2b switch variants of the mAb did not prevent infection of the genital tract but resulted in lower virus loads in the vaginal epithelium and provided significant protection against disease and acute infection of the sensory ganglia independently of host FcγR expression. Together, these data demonstrate two distinct antibody effector mechanisms capable of providing substantial protection against genital HSV-2 disease. The presence of either FcγR -dependent Ab or strongly neutralizing Ab did not completely prevent HSV-2 infection but limited initial infection of genital and neuronal tissues and diminished HSV-2 disease. NIH Grants AI42815 and AI054444.
Oral tolerance defects in congenital T cell-deficient mice (42.1)
Authors: Lee Joon Youb , Oh Nicholas , Surh Charles D 
Published: January 2023

Abstract
Congenital T cell (T)-deficient mice, such as RAG− or SCID mice, succumb to inflammatory bowel disease (IBD) upon adoptive transfer of syngenic naïve T cells - a consequence of chronic inflammation in the intestine in response to commensal bacterial antigens. Currently, the underlying reason for the onset of IBD is believed to be the absence of regulatory T cells (Tregs). However, since donor T cells undergo considerable chronic activation even in the presence of Tregs, abnormalities in the presentation of enteric antigens also appears to be involved. To further explore this idea, we have compared the fate of TCR transgenic T cells adoptively transferred into normal vs. RAG− mice that are fed with the cognate antigen. Strikingly, while the donor TCR transgenic T cells undergo abortive slow cell division in normal hosts and slowly disappear, the same cells undergo rapid proliferation, differentiate into effector cells and persist for prolonged periods. These findings suggest that RAG− mice have an inherent defect in their ability to induce oral tolerance.
Exacerbation of spontaneous colitis in the absence of flora derived TLR4 signals (42.2)
Authors: Mingle Bethany , Afshar Roshi , Prioult Guenolee , Murakami Hidehiro , Nagler Cathryn 
Published: January 2023

Abstract
Complex immunoregulatory mechanisms are required to maintain non-responsiveness to dietary antigens and the commensal flora. We have reported that mice that are unable to signal via TLR4, the receptor for bacterial LPS, are highly susceptible to an allergic response to food. When the composition of the commensal flora is reduced and altered by antibiotic treatment, TLR4 wild-type mice become as susceptible to the induction of allergy as their TLR4 mutant counterparts. More recently we have shown that the mesenteric lymph nodes of TLR4 mutant mice are deficient in the generation of IL-10 secreting regulatory T cells (Tregs) and lack a population of plasmacytoid DC that constitutively makes IL-10. To further examine the contributions of IL-10 secreting Tregs and DC to mucosal non-responsiveness, we prepared TLR4xIL-10 double knockout (DKO) mice. We have found that the Th1 biased colitis previously reported in IL-10 deficient mice is greatly exacerbated in TLR4xIL-10 DKO mice. Our observations stand in contrast to a recent report from Rakoff-Nahoum et al (Immunity 2006 25:319) that shows that MyD88xIL-10 DKO mice are protected from the development of colitis. TLR4 is unique among bacteria responsive TLRs in its ability to signal via both MyD88 dependent and MyD88 independent pathways. Taken together with our observation that food allergy is linked to an inability to signal via TLR4, the exacerbation of spontaneous colitis in TLR4xIL-10 DKO mice supports a unique role for TLR4 signaling in the generation of flora induced immunoregulatory cells that control pathological responses at both ends of the Th1/Th2 spectrum.
Supported by NIH grants DK 55678 and DK43351.
CD8aa-specific CD8ab Effector Memory T Cell Differentiation in the Gut (42.6)
Authors: HUANG YUJUN , Schoenberger Stephen , Cheroutre Hilde 
Published: January 2023

Abstract
CD8 effector memory T cells at the intestinal mucosa are important for rapid containment of the invading pathogens. CD8aa, a homodimer of CD8a, has been found on gut intraepithelial lymphocytes (IELs). It is also transiently expressed on some peripheral CD8ab primary effector T cells that differentiate to memory T cells. In vitro stimulated sorted CD8aa+ and CD8aa− OT-I cells migrate with equal capacity to the gut and spleen of recipient mice. One month later, oral re-challenge with Listeria Monocytogenes-OVA (LM-OVA) leads to an immediate secondary expansion of CD8aa+, but not CD8aa−, donor cells among the IELs, which suggests that only CD8aa+ precursors can differentiate into mucosal memory T cells and respond immediately to an oral pathogen challenge. Later, both CD8aa+ and CD8aa− donor cell-derived secondary responses are detected in spleen and gut. However, 45 days after the secondary response, only CD8aa+donor cells generate memory IELs. It indicates that CD8aa+ primary effector cells are programmed during the initial priming to survive and to become mucosal effector memory T cells. CD8aa expression is transient on primary effector cells and regained after relocation into the gut. The specific role of CD8aa in the survival and differentiation of intestinal CD8 effector memory T cells was investigated.
Influenza specific CD8 T cells require interactions with pulmonary DC in the lungs in order to mount an effective immune response (42.8)
Authors: McGill Jodi , Legge Kevin 
Published: January 2023

Abstract
Our studies show that respiratory dendritic cells (rDC) transiently migrate (i.e. within the first 48 hours post infection) from the lungs to the lymph nodes (LN) following influenza virus infections. However, DC are continually recruited into the lungs even after DC migration to the LN has ceased. Since CD8 T cell activation and programming occurs in the LN, the role of these lung-recruited DC in influenza immunity was unclear. To better understand their role, we depleted rDC after the conclusion of normal rDC trafficking to the LN and assessed the immune response. Our results show decreased CD8 T cell responses, resulting in increased viral titers, morbidity and mortality. Initial T cell expansion and activation in the LN, and migration to the lungs appears equivalent in both wildtype and rAPC depleted mice (rAPCneg). However, in rAPCneg mice, these activated T cells fail to continue their expansion once arriving in the lungs. Examination of the DC subtypes present in the lungs revealed decreased numbers of both pulmonary pDC and CD8+ DC in rAPCneg mice through 6 days p.i. Reconstitution of rAPCneg mice with purified pulmonary pDC or CD8+ DC restores influenza-specific CD8 T cell immunity in a MHC I dependent manner. These results suggest that activated influenza-specific CD8 T cells may require additional antigen-specific interactions with recruited DC in the lungs in order to protect against influenza virus infections.
Deterministic linkage between CD4 T cell and B cell specificities to a large virus: vaccinia virus (43.1)
Authors: Crotty Shane , Moutaftsi Magdalini , McCausland Megan , Davies D. Huw , Quiroz Juan Moyron , Johnston Robert , Benhnia Rafii Mohammed , Grey Howard , Hoffmann Julia , Head Steven , Garboczi David , Sette Alessandro 
Published: January 2023

Abstract
Antibody responses are critical components of protective immune responses to many pathogens, but it remains unclear what parameters determine which pathogen proteins are targeted. We have performed large scale studies to map mouse antibody and CD4 T cell responses to the smallpox vaccine virus, vaccinia. Our analysis of this large virus ( &gt; 200 genes) identified 13 viral protein targets of CD4 T cell responses and 23 antibody targets. Unexpectedly, nearly half of the CD4 T cell responses targeted the same viral virion proteins as the antibody responses, suggesting a deterministic linkage between the specificities. Immunizing mice with individual MHC II epitopes revealed that, after viral infection, help between CD4 T cells and B cells of matched viral virion protein specificity is largely non-transferrable to other virion proteins. We used this knowledge to then predict the presence of new MHC II epitopes. In total, 11 of the 18 CD4 T cell responses (61%) we identified were matched by a paired antibody response to the same protein, including all of the top 5 viral protein IgG targets. These data demonstrate that MHC restriction at the protein level is a critical event for anti-virion antibodies to a large virus. This is a powerful predictive principle and is of general relevance for understanding the nature of B cell antigen presentation to CD4 T cells against complex pathogens. NIH-AI-63107
Chlamydia pneumoniae infection modulates cytokine production by human T lymphocytes and monocytes (43.11)
Authors: Hakki Amal , Mamata Yukimitsu , Friedman Herman 
Published: January 2023

Abstract
Continuous cell line cultures of human lymphocytes (MOLT-4) and monocytes (THP-1) were examined for production of immunoregulatory cytokines involved in immune responses to the intracellular opportunistic pathogen Chlamydia pneumoniae (Cp). Infection with Cp is widely prevalent and believed involved in chronic inflammatory diseases such as atherosclerosis and even neurodegenerative diseases like Alzheimer’s disease and multiple sclerosis. The results of this study showed that MOLT-4 and THP-1 cells were readily infected by Cp in vitro assessed by immunofluorescence microscopy for Cp lipopolysaccharide (LPS) and by real time RT-PCR for 16S rRNA expression. Stimulation of MOLT-4 cells with heat killed Cp (KCp) or Escherichia coli LPS (E. coli LPS) induced Th1 cytokines IFNγ and IL-12 and Th2 cytokine IL-10, but infection with viable Cp showed a dominant Th1 cytokine production. Cp infected THP-1 cells mainly produced IL-12 and enhanced TNFα and IL-10 production compared to cultures stimulated with KCp or E. coli LPS as a control. These results suggest that Cp infection modulates the immune response to a Th1 biased cytokine profile by T cells in addition to induction of TNFα by monocytes/macrophages. Such effects are likely involved in antibacterial immunity against Cp infection.
Phenotype and functional properties of Vaccinia specific CD8+ T cells (43.12)
Authors: Ostrout Nicholas D , McHugh Marilyn M , Brustoski Kim , Kazura James 
Published: January 2023

Abstract
Memory CD8+ T cells play a significant role in mediating protective immunity from pathogens, particularly viruses and intracellular bacteria. Inducing the optimal response, from both a quantitative and qualitative perspective, has been a research priority for determining the effectiveness of new vaccines. Depending on the pathogen, central memory (TCM) or effector memory (TEM) may be more important in providing adequate protection. Therefore, when developing new vaccines, it is important to realize which memory CD8+ T cell subset is vital and potentially ‘drive’ the response to elicit a protective level ratio in favor of one subset. Here we describe the phenotype of vaccinia specific memory CD8+ T cells. TEMRA+ memory T cells produce the most IFN-γ of any memory subset and are present in individuals last vaccinated over 30 years ago. Concurrently, we address the development of memory CD8+ T cells upon administration of Dryvax, as well as new smallpox vaccines such as Modified Vaccinia Ankara (MVA) in a murine model. Previous studies have indicated that TEM appear to provide better secondary protection than TCM and the data presented here indicate that these cells are maintained long term (&gt;30 years). Therefore, we propose that new smallpox vaccines, such as MVA, should elicit a memory cell population in favor of TEM similar to that induced by Dryvax or other traditional smallpox vaccines.
cAMP response element binding protein (CREB), activating transcription factor (ATF)-2 and c-Jun bind to the IFN-γ proximal promoter in response to M. tuberculosis (TB) stimulation (43.13)
Authors: Samten Buka , Townsend James C. , Weis Stephen E. , Barnes Peter F. 
Published: January 2023

Abstract
The proximal promoter controls transcription of IFN-γ in T cells, which is central for defense against many pathogens. By performing EMSA and supershift assays on nuclear extracts of TB-stimulated PBMC, we found that CREB, cAMP response element modulator (CREM), ATF-1, ATF-2, and c-Jun bound to the IFN-γ proximal promoter. Microaffinity purification with a biotinylated IFN-γ proximal promoter, followed by Western blotting, showed that only CREB, ATF-2 and c-Jun bound to the proximal promoter, and this was confirmed in live T cells by chromatin immunoprecipitation. Coimmunoprecipitation of nuclear extracts of TB-stimulated PBMC indicated interaction of these transcription factors. Introduction of a plasmid expressing a dominant-negative ATF-2 peptide into TB-stimulated T-cells inhibited IFN-γ secretion by &gt;90% and reduced expression of CREB, ATF-2 and c-Jun. Nucleofection of siRNA specific for ATF-2 decreased expression of CREB, c-Jun and ATF-2 in T cells. TB-stimulated PBMC from tuberculosis patients, known to have reduced IFN-γ production, were found to have low levels of CREB, ATF-2 and c-Jun, but not CREM, ATF-1 and c-Fos, compared to PBMC from healthy donors. We conclude that:
CREB, ATF-2 and c-Jun bind to the IFN-γ proximal promoter;ATF-2 contributes to expression of CREB and c-Jun;CREB, ATF-2 and c-Jun contribute to T-cell production of IFN-γ in response to TB.
(NIH AI063514)
CXCL10 is required for the recruitment of leukocytes responsible for controlling herpes simplex virus-1 infection (43.19)
Authors: Wuest Todd , Luster Andrew , Campbell Iain , Farber Joshua , Carr Daniel 
Published: January 2023

Abstract
The chemokine CXCL10 is among the earliest cytokines produced during nervous system inflammation induced by herpes simplex virus-1 (HSV-1) infection. Current evidence suggests CXCL10 is important in the recruitment of a wide range of leukocytes including CD4 and CD8+ T-cells, macrophages, dendritic, and NK cells. Many of these studies have suggested the function of this chemokine extends beyond simple leukocyte recruitment as CXCL10 is also expressed in primary and secondary lymphoid tissues. To elucidate the role CXCL10 has in generating an effective immune response to viral infection of the nervous system, we have analyzed leukocyte recruitment and effector function in CXCL10 deficient (−/−) mice relative to wild type C57BL6/J, CXCL9 deficient, and CXCR3 deficient mice infected with HSV-1. Our results indicate CXCL10 is critical for the recruitment of NK cells and B220+CD11c+ cells, in a CXCR3-dependent manner. Infiltration of HSV-1 specific CD8+ T-cells into the central nervous system of CXCL10 −/− mice is significantly reduced compared to wild type mice, but CXCL10 expression in lymphoid tissues does not contribute in the generation of HSV-specific CD8+ T-cells. Nervous system restricted expression of CXCL10 restored the homing of HSV-1 specific CD8+ T-cells to wild type levels and partially restored homing of natural killer and conventional dendritic (B220-CD11c+) cells.
Transgenic expression of Bcl-xL or Bcl-2 by murine B cells enhances the in vivo anti-polysaccharide, but not anti-protein, response to Streptococcus pneumoniae (43.24)
Authors: Chattopadhyay Gouri , Khan Abdul Q. , Dubois Wendy , Potter Michael , Snapper Clifford M. 
Published: January 2023

Abstract
IgG anti-polysaccharide (PS) and anti-protein responses to Streptococcus pneumoniae (Pn) are dependent on CD4+ T cells, B7/CD28, and CD40/CD40-ligand interactions. However, the primary IgG anti-PS response terminates more quickly, requires a shorter period of T cell help, fails to generate memory, and is more dependent on Ig signaling. To determine a role for B cell apoptosis in these differences, we utilized mice expressing an Ig heavy chain enhancer-driven transgene encoding the anti-apoptotic proteins Bcl-xL or Bcl-2. Bcl-xL-Tg and Bcl-2-Tg mice immunized with Pn exhibited a prolongation of the primary IgM and IgG anti-PS, but not anti-protein, response resulting in up to 10-fold enhancements in specific Ig serum titers. Of note, Tg mice still failed to generate PS-specific memory. In contrast, Fas- or Fas-ligand-deficient mice elicited normal humoral immune responses to Pn. In vitro, Bcl-xL and Bcl-2 Tg B cells exhibited higher and more prolonged proliferative responses following multivalent Ig crosslinking, but not after stimulation with LPS or anti-CD40 mAb, relative to WT B cells. In contrast to intact Pn, in vivo anti-PS responses to purified PS or a soluble PS-protein conjugate were similar in Tg and WT mice. These data suggest that apoptosis may selectively limit clonal expansion and subsequent Ig secretory responses of PS-specific B cells during a primary immune response to an intact bacterium.
N.I.H. R01 AI49192
Differential role of CD4 and CD8 T cells in RSV immune pathology (43.28)
Authors: Neupane Rupak , Schaller Matthew , Lukacs Nicholas W 
Published: January 2023

Abstract
CD4 and CD8 T cells produce different cytokines during immune response. Using a model of respiratory syncytial virus (RSV) infection, isolated CD4 T cells were found to produce predominantly Th2, while CD8 produced predominantly Th1 cytokines. To better understand the contribution of CD4 and CD8 T cells in immune pathology during RSV infection, CD4 and CD8 T cells were depleted using specific depleting antibodies and immune response was compared with control mice during RSV infection. Depletion of CD8 T cells showed increased airway hyperresponsiveness (AHR), production of IL-13, as well as increased mucus-associated gene, Gob5, expression. Specific depletion of CD4 induced lower AHR compared to control mice, lower Th2 cytokines and reduced Gob5 production. PAS staining showed both control and CD8 depleted mice with increase mucus production and increased goblet cell hyperplasia whereas mice depleted of CD4 T cell did not produce mucus. These results indicate that CD8 T cells are protective whereas CD4 T cells are responsible for increased pathology observed during RSV infection.
This work was supported in part by NIH RO1AI036302.