bluuebunny/crossref_metadata_2025_split huggingface dataset:
Memory CD8+ T cells deficient in IFN-γ protect from a lethal viral disease (105.3)
Authors: Remakus Sanda , Rubio Daniel , Sigal Luis 
Published: January 2023

Abstract
Ectromelia virus (ECTV), the causative agent of mousepox, is a lethal mouse pathogen of the Orthopoxvirus genus, which also includes variola virus. C57Bl/6 (B6) mice are resistant to ECTV infection while mice such as DBA2/J are susceptible. B6.D2-D6 mice, a B6 congenic strain with the NK complex from DBA2/J are sensitive to ECTV and can be protected by adoptive transfer of memory CD8+ T cells (MTCD8+) from convalescent mice. To ascertain the role of IFNγ in MTCD8+ mediated protection, we transferred MTCD8+ from IFNγ-/- mice (Mγ-/-) into B6.D2-D6 mice and challenged them with ECTV. All naïve CD8+ (NB6) recipients died and all recipients of memory B6 CD8+ (MB6) or Mγ-/- recipients survived. Seven dpi, MB6 and Mγ-/- recipients had significantly less liver damage and virus titers in the spleen and liver than NB6 recipients. This demonstrates that MTCD8+ do not have to produce IFNγ to protect from a viral disease. Of interest, we also found that donor virus-specific cells promoted an endogenous virus-specific TCD8+ response in the spleens and livers of recipients. Next we determined if IFNγ was necessary when MTCD8+ cells were present by transferring MB6 or Mγ-/- CD8+ into IFNγ-/- mice. All NB6 and Mγ-/- recipients succumbed to mousepox while all recipients of MB6 recipients survived. Thus, IFNγ is required for resistance to mousepox but can be produced in sufficient quantities for protection by MTCD8+ or other non-memory cells, probably NK cells or primary T cell effectors.
Comprehensive evaluation of the CD8 responses to HSV-1 in humans (105.41)
Authors: Jing Lichen , Haas Juergen , Dann Greg , Dong Lichun , Liang Kerry , Wald Ann , Koelle David 
Published: January 2023

Abstract
CD8-only vaccines can be protective on some mouse models. HSV-1-specific CD8s localize to sites of lesion and healed skin, and to chronically infected ganglion. We enriched polyclonal HSV-1-specific CD8s from human blood using a novel cross-presentation/activation marker-based protocol. After expansion, these cell populations were quite oligoclonal as assessed by TCR beta chain CDR3 deep sequencing. To query fine specificity, artificial APC were created by transfecting Cos-7 cells with each HLA A or B heavy chain from the subject. Each HSV-1 ORF was cloned into an expression vector and separately co-transfected with each HLA allele. Activation of CD8+ cells in the polyclonal T-cell population to specific ORFs was detected by interferon-gamma ELISA. The complexity of the response ranged from 7 to 23 reactive ORF/HLA combinations per subject. Most such epitopes are HSV-1 type-specific, but some are identical within the related pathogen HSV-2. One type-common epitope was studied in HLA A*0201-transgenic mice, in which DNA vaccination with the HSV-1 full-length ORF lead to very strong epitope-specific responses. Overall, we observed a remarkably similar complexity for the CD8 response to HSV-1 by using either the deep TCR sequencing or antigenic specificity method. Novel candidate immunodominant HSV-1 CD8 antigens were identified, and no apparent skewing to virion input or immediate early proteins due to TAP inhibition was detected.
The tug-of-war between dendritic cells and human chronic viruses (105.47)
Authors: Jain Pooja 
Published: January 2023

Abstract
The human immune system is under constant challenge from many viruses, some of which the body is successfully able to clear. Other viruses have evolved to escape the host immune responses and thus persist, leading to the development of chronic diseases. Dendritic cells (DCs) are professional antigen presenting cells that play a major role in both innate and adaptive immunity against different pathogens. For the past few years our efforts have been focused on exploring the participation of DCs in the pathogenesis of HTLV-1, HIV-1 and HCV. We observed previously that depletion of DCs in CD11c-DTR transgenic mice enhanced the susceptibility to cell-free HTLV-1 infection. We further performed the host-pathogen interaction studies utilizing Flt3 ligand derived murine bone marrow DCs (FL-DCs). First, the kinetics of viral entry, proviral integration, and expression of the viral protein Tax was established and then effects of cell-free HTLV-1 was examined on these cells. Phenotypically, FL-DCs demonstrated activation and produced an array of proinflammatory cytokines as well as IFN-α. Virus-matured FL-DCs also stimulated proliferation of autologous CD3+ T cells and IFN-γ production. Gene expression studies revealed upregulation of interferon-stimulated genes, most cytokines, and transcription factors but a distinct downregulation of many chemokines. Overall, these results highlight the critical interaction of DCs with a human chronic virus important for the early immune responses.
A novel, protective Toll-like receptor independent innate immune response to HSV-1 in the cornea (105.51)
Authors: Conrady Christopher , Zheng Min , Wuest Todd , Mandal Nawajes , Tran Julie , Liu Chuanju , Carr Daniel 
Published: January 2023

Abstract
Toll-like receptors (TLRs) are innate sentinels required in clearance of bacterial and fungal infections of the cornea, but their role in viral immunity is currently ambiguous. To evaluate the role of TLRs in ocular immunity to viral infection, TLR adaptor protein deficient mice (MyD88-/- and Trif-/-) were infected with HSV-1. MyD88-/- and Trif-/- mice had similar viral loads in the cornea as those of wild type (WT) mice but substantially lower levels than type 1 interferon receptor A1 deficient (CD118-/-) mice 48-120 hours post infection (pi). A chimeric mouse model utilizing WT and CD118-/- mice was then employed and identified both a resident and bone marrow (BM)-derived immune component in viral containment 48 hours pi. The BM contribution was linked to infiltrating F4/80+ GR1+ monocytes that were severely diminished in CD118-/- mice compared to all other groups, corresponding to a loss of CCL2 production. CCL2 deficient mice were found to possess significantly more virus, consistent with a loss of F8/80+ GR1+ infiltrating monocytes in the cornea compared to WT controls. To identify the key sentinel in viral recognition of the cornea, the DNA sensor, p204, was first identified in mouse corneal epithelium by confocal microscopy and Western blot. In vivo siRNA knockdown of the sensor resulted in significant elevation of virus. Taken together, these results describe a novel TLR-independent innate immune sensor critical in the initial control of acute HSV-1 infection.
Nanolipoprotein particles as vaccine platforms for co-localization of adjuvants and subunit antigens (106.10)
Authors: Blanchette Craig , Hoeprich Paul , Fischer Nicholas 
Published: January 2023

Abstract
Subunit vaccines provide a number of important benefits over traditional vaccine candidates, such as overall safety. However, these antigens require immunostimulatory adjuvants to generate the appropriate adaptive immune response. We hypothesize that the efficacy of subunit antigens will be enhanced if co-localized with adjuvant during delivery in a single delivery platform. Thus, our goal is to develop a robust vaccine delivery platform that will allow for the co-delivery of both subunit antigens and adjuvants using nanolipoprotein particles (NLPs). NLPs are discoidal HDL analogs comprised of lipid bilayers stabilized by scaffold proteins. These biocompatible, nanoscale particles can be assembled to incorporate nickel-chelating lipids to form nickel-chelating NLPs (NiNLPs). Thus, any His-tagged antigens can be conjugated to the NiNLPs. The amphipathic adjuvants monophosphoryl Lipid A (MPLA) and cholesterol-modified CpGs (Chol-CpGs) were anchored to the lipid bilayer of the NiNLPs. His-tagged influenza hemagglutinin 5 was conjugated to adjuvant:NiNLPs and injected into mice. Antibody titers increased 32 and 8 fold (MPLA and CpG, respectively) compared to co-administered formulations. Also, the immune response was skewed towards a Th1 response, with up to 16-fold increases in IgG2a:IgG1 ratios. Our findings demonstrate that co-localized delivery of adjuvant with antigens provides significantly greater immune stimulation in mice compared to co-administered formulations.
Dendritic cell function and T cell responses to influenza viruses in patients with X-linked agammaglobulinemia (106.12)
Authors: Liu Yinping , Yuet Wu , Tu Wenwei , Lau Yu-Lung 
Published: January 2023

Abstract
Immunodeficiency patients have high risks for influenza infection. X-linked agammaglobulinemia (XLA) is a primary immunodeficiency characterized by the deficiency of humoral immunity. Hence, cellular immune response becomes more important for these patients. However, little is known about the non-humoral immune responses to influenza virus in XLA patients. Here, we examined the APC function and T-cell immune responses to influenza viruses in XLA patients. Monocyte-derived immature dendritic cells (imMDDCs) from XLA patients had normal maturation upon inactivated influenza virus stimulation in terms of surface molecule expressions and cytokine productions. The T-cell stimulatory capacity of influenza virus-treated imMDDCs determined by mixed lymphocyte reactions from XLA patients was also similar as that from normal controls. There were no obvious side effects in XLA patients after administration of trivalent inactivated influenza vaccine (TIV). TIV significantly induced influenza virus-specific CD4 and CD8 T-cell responses in XLA patients. The frequencies of influenza virus-specific memory/effector CD4 and CD8 T cells in XLA patients are comparable to that in normal controls during 6 months of observation after vaccination. Our results indicated that XLA patients have normal APC function and T cell responses to influenza virus. Influenza vaccination may provide some benefits to XLA patients for influenza infection, at least reducing the severity of illness.
Control of MRSA sepsis by CCL1 antisense ODN in severely burned mice (106.23)
Authors: Asai Akira , Kogiso Mari , Herndon David , Kobayashi Makiko , Suzuki Fujio 
Published: January 2023

Abstract
Severely burned mice are very susceptible to MRSA infection. M1Mϕ, a major effector cell in host antibacterial innate immunity, were not generated in mice 1 to 3 weeks after severe burn injury, because these mice carried M2bMϕ predominantly. Previously, CCL1 was shown to be needed to maintain M2bMϕ properties, and the elimination of M2bMϕ was demonstrated in burned mice treated with CCL1 antisense ODN. Therefore, in this study, we tried to control MRSA infection by CCL1 antisense ODN treatment in mice 1 to 3 weeks after burn injury. Burned mice (25% TBSA, 3rd degree flame burn) were treated s.c. with 10 μg/mouse of CCL1 antisense ODN twice a day for 2 days beginning 13 days after burn injury. Fourteen days after burn injury, these mice were infected i.v. with 107 CFU/mouse of MRSA. The severities of MRSA infection were evaluated by (i) the growth of the pathogen in organs and blood and (ii) their mortality rates, as compared with controls (burned mice treated with scrambled ODN). In the results, all burned mice treated with CCL1 antisense ODN survived after MRSA infection, while 83% of burned mice treated with scrambled ODN died. After infection, MRSA grew in the kidneys and blood of burned mice treated with scrambled ODN, while greatly diminished growth of the pathogen was shown in these organs of burned mice treated with CCL1 antisense ODN. These results indicate that MRSA sepsis is effectively controlled in burned mice subjected to CCL1 gene therapy.
Anti-lipopolysaccharide IgM antibody exhibits vibriocidal activity against Vibrio cholerae in the clinical sera (106.4)
Authors: Yang Jae Seung , Kang Seok-Seong , Kim Kyoung Whun , Yun Cheol-Heui , Han Seung Hyun 
Published: January 2023

Abstract
Serum vibriocidal antibody assay has been used to evaluate the immunogenicity of cholera vaccines formulated with killed whole-cell of Vibrio cholerae. However, serum antibodies representing the vibriocidal activity are not fully characterized. In this study, we examined which isotype is associated with the vibriocidal activity using human convalescent sera with high virbiocidal activity and 20 clinical serum samples obtained from human subjects who had been vaccinated with a killed, whole-cell cholera vaccine. Total IgG, IgA or IgM antibody levels were not correlated with the vibriocidal activity in the 20 human clinical sera. When IgG, IgA and IgM antibodies specific to each of the heat-killed V. cholerae, lipopolysaccharide (LPS), or outer membrane protein U were measured in the 20 clinical serum samples, anti-LPS IgM antibody showed the highest correlation with the vibriocidal activity (Pearson correlation coefficient r = 0.818). Depletion of IgM antibodies in the convalescent sera produced a significant (P&lt;0.05) decrease in the vibriocidal activity (16-fold decrease), whereas little change was observed with depletion of IgG and/or IgA. Furthermore, size-exclusion column chromatography followed by vibriocidal assay and analysis of antibody isotypes demonstrated the co-elution of vibriocidal activity and IgM antibody. Collectively, these results suggest that LPS-specific IgM could be a potential serologic marker for the vibriocidal antibody assay.
Anti-CD3 mAb therapy promotes tolerance by selectively depleting pathogenic cells while preserving regulatory T cells (107.12)
Authors: Penaranda Cristina , Tang Qizhi , Bluestone Jeffrey 
Published: January 2023

Abstract
Monoclonal anti-CD3 antibodies have been used clinically for two decades to reverse steroid-resistant acute graft rejection. In autoimmune diabetes, short course treatment with FcR non-binding anti-CD3 mAb in mice with recent onset of diabetes induces long-term disease remission. Induction of tolerogenic regulatory T cells (Tregs) has been implicated to be one of the mechanisms of action by FcR non-binding (FNB) anti-CD3 mAb in these settings. In this study, we examined the effect of FNB anti-CD3 mAb treatment on the homeostasis of CD4+CD25+Foxp3+ Treg cells in vivo. Anti-CD3 mAb treatment induced a transient systemic rise in the percentage but not absolute number of Tregs due to selective deletion of CD4+CD25- naïve and effector T cells. T cell depletion induced by FNB anti-CD3 mAb was independent of pro-apoptotic proteins Fas or Bim and was caspase 3 independent. Tregs were not preferentially expanded and we found no evidence of convertion of conventional T cells into Tregs suggesting that Tregs are resistant to anti-CD3-induced cell death. Interestingly, we found increased Helios expression on Tregs further showing that adaptive Tregs are not induced by anti-CD3 treatment. Taken together, our results suggest that FNB anti-CD3 mAb therapy selectively induce death of potentially pathogenic CD25- T cells and stabilized, although did not induce, Tregs, thus promoting tolerance by restoring the balance between pathogenic T cells and regulatory T cells.
Structural requirements for the inhibition of calcium mobilization and mast cell activation by the pyrazole derivative BTP2 (107.16)
Authors: Law Mankit , Mottram Laurie , Morales Jose , Peterson Blake , August Avery 
Published: January 2023

Abstract
Cytosolic Ca2+ changes in mast cells are central for driving their activation. Thus, pharmacological tools that inhibit this process represent unique strategies for inhibiting mast cell function and their contribution to disease. We find that the pyrazole compound BTP2 inhibited FcϵRI-triggered sustained Ca2+ influx in RBL-2H3 cells and murine BMMCs with little effect on the tyrosine phosphorylation of cellular proteins. It also did not affect FcϵRI-induced phosphorylation of mitogen-activated protein kinases Erk1/2, JNK, and p38 nor downstream c-fos expression. This suggests that BTP2 inhibits Ca2+ mobilization without affecting the FcϵRI signal transduction cascade. In vitro, BTP2 inhibited FcϵRI-triggered degranulation and cytokine secretion, correlating with inhibition of NFAT nuclear translocation. Consistent with this effect, BTP2 inhibited antigen-induced histamine release in vivo. Analysis of structure-function relationships between the BTP2 parent compound and its derivatives implicate a role for the trifluoromethyl group at both carbon positions 3 and 5 of pyrazole in the inhibition of degranulation of BMMCs, with a more dominant effect evoked by the trifluoromethyl group at C3. In conclusion, our studies indicate that pyrazole compounds can provide the molecular framework for the design of specific inhibitors of Ca2+ entry mechanisms and further BTP2 as a medicinal candidate for the treatment of allergic reactions and other mast cell-mediated diseases.
The effect of B-cell depletion in Theiler's viral model of multiple sclerosis (107.17)
Authors: Li Libin , Matsumoto Makiko , Pachner Andrew 
Published: January 2023

Abstract
Introduction: B cell depletion (BCD) has been utilized to treat relapsing-remitting multiple sclerosis (MS), but it is also not clear whether BCD ameliorates disability progression. We used an anti-mouse CD20 mAb (5D2), and tested its effects in Theiler’s murine encephalomyelitis virus-induced demyelinating disease (TIDD), a viral CNS infection considered the best experimental animal model of disability progression in MS. Methods: SJL mice were injected intraperitoneally with 5D2 15 days before intracerebral infection of TMEV BeAn strain. 5D2 bioactivity was assessed by flow cytometry assay of CD19+ cells in peripheral blood mononuclear cells (PBMC). Mouse disability was measured by Rotarod, and viral load was measured by real-time reverse transcriptase PCR (RT-PCR) for TMEV RNA. Antibody levels were determined in mouse sera and CSF by ELISA, and in CNS tissues by real-time RT-PCR for IgG RNA. Results: Bioactivity of 5D2 was confirmed by nearly absent CD19+ cells in the peripheral blood from animals with persistent BCD. Antibody responses in both CNS and peripheral system were suppressed during 5D2 treatment. Relatively higher viral loads were detected in 5D2-treated mice than in PBS animals, and the viral levels correlated negatively to IgG production in the brain. 5D2 caused marked worsening of the early encephalitis, possibly from deletion of a B cell phenotype which downregulates inflammation.
Combined antibody therapy for type 1 diabetes (107.2)
Authors: Hu Changyun , Ding Heyuan , Wong F. Susan , Wen Li 
Published: January 2023

Abstract
Studies have shown that oral tolerance is effective in treating different models of autoimmune diseases including type 1 diabetes (T1D). Both T and B cells play important roles in the development of T1D. Antibodies targeting either T or B cells have shown to be effective in treating T1D in both patients and NOD mice. However, individual antibodies are only effective in a proportion of those treated. Furthermore, the beneficial effects have not been long lasting. To test whether the combination of anti-CD20 and oral anti-CD3 treatment can improve the efficacy in prevention and reversal of T1D and prolong the beneficial effects, we treated a group of human CD20 transgenic NOD mice with anti-hCD20 monoclonal antibody and oral anti-CD3. The results suggested that anti-CD20 and anti-CD3 have synergistic effects in the diabetes prevention and reversal. Mechanistic studies have demonstrated that the combination treatment induced FoxP3+ Tregs and enhanced the regulatory function of Treg. A subset of IL-10+ CD4 T cells was also induced in peyer’s patches of the mice treated with combined therapy. Our preliminary study may provide a novel therapeutic approach to treating T1D.
Development of antigen-loaded, immunosuppressive microparticles to modify dendritic cell behavior for the prevention of type 1 diabetes in NOD mice. (107.9)
Authors: Keselowsky Benjamin , Lewis Jamal , Carstens Matthew , Xia Chang Qing , Clare-Salzler Michael 
Published: January 2023

Abstract
Antigen-specific immunomodulation remains an important goal in the treatment of type 1 diabetes (T1D). We hypothesize that in vivo targeting of biodegradable microparticles (MPs) to DCs delivering immunomodulatory agents can promote a tolerogenic DC phenotype and induce Tregs in vivo, in order to ameliorate type 1 diabetes in NOD mice. To test this hypothesis, poly(lactide-co-glycolide) MPs were loaded with antigen - insulin B:9-23, and immunomodulatory factors - vitamin D3 (D3), TGF-B1 and GM-CSF. These MP formulations were injected subcutaneously into 4 week old female NOD mice followed by a booster at 5 weeks of age. Formulations (n = 10) consisted of a.) blank MPs, b.) MPs loaded with insulin peptide only, c.) MPs loaded with GM-CSF + insulin peptide, d.) MPs loaded with D3+TGF-B1+insulin peptide, e.) MPs loaded with GM-CSF+D3+TGF-B1 + insulin peptide. Blood glucose was measured once a week until 32 weeks of age (currently at 22 weeks). Kaplan-Meier analysis at this point reveals an overall p-value of 0.094 for survival proportions among groups, with the highest proportion of non-diabetic mice attributed to the formulation of MPs loaded with GM-CSF+D3+TGF-B1+insulin peptide (60% non-diabetic), compared to the blank MPs with the lowest proportion (20% non-diabetic), suggesting delayed onset of T1D. In vitro mechanistic assessments demonstrate MP formulations are capable of promoting tolerogenic DC phenotype and inducing syngeneic CD4+CD25+foxp3+ regulatory Tcells.
Immunity to secondary bacterial pneumonia caused by Streptococcus pneumoniae following Influenza A infection (108.10)
Authors: Bernui Mariana , Liboon Jennie , Shen Hao 
Published: January 2023

Abstract
IAV is highly contagious and responsible for both seasonal and pandemic flu. However, mortality associated with IAV is mainly due to secondary bacterial infections. The bacterial and host factors that mediate lethality, as well as the protective immune mechanisms against co-infections, are largely unknown. By sequentially infecting mice with IAV and Sp (IAV/Sp), we show that IAV-infected mice are highly susceptible to secondary Sp infections (&lt;10 CFU). Mice lacking IL-12, Type 1 IFN receptors, or both were just as susceptible to an IAV/Sp infection, suggesting these inflammatory cytokines are not involved in lethality. IAV-specific CD8 T cells from IAV/Sp co-infected mice had increased PD-1 expression, a marker associated with dysfunctional/exhausted CD8 T cells. When we immunized mice with different strains of IAV or Sp and challenged them with IAV/Sp co-infection, IAV-immunized mice were fully protected against an IAV/Sp infection. Immunization with one IAV strain provided cross-protection against a co-infection challenge with a different serotype strain of IAV. Immunization with Sp prolonged survival of co-infected mice, and was IL-12 dependent. We conclude that 1) Early inflammatory cytokines are not responsible for death caused by IAV/Sp co-infections, 2) Secondary Sp infections do not affect the magnitude but the quality of IAV-specific CD8 T cell responses, and 3) cross-protective immunity against IAV/Sp co-infections can be induced by immunization with IAV or Sp.