bluuebunny/crossref_metadata_2025_split huggingface dataset:
The immunotherapeutic GEN-003 as a prophylactic vaccine candidate for genital herpes in guinea pigs
Authors: Kaufmann Johanna K , Cardin Rhonda D , O’Keeffe Michael , Baccari Amy , Le Bonnie , Sanz Vinny , Yu David , Flechtner Jessica B , Flano Emilio , Bernstein David I 
Published: January 2023

Abstract
GEN‑003 is a subunit vaccine candidate containing adjuvanted gD and ICP4 antigens that has shown durable therapeutic efficacy against genital herpes in a phase 2 clinical trial. Here we describe the potential benefit of GEN-003 as a prophylactic vaccine in a guinea pig model of genital herpes.
Animals were immunized three times every two weeks with GEN-003 or control and intravaginally challenged with herpes simplex virus 2 (HSV-2, MS strain) 21 days later. GEN-003 significantly reduced frequency and severity of genital lesions in both acute and early stages of recurrent infection by over 85%, and 60% of vaccinated guinea pigs did not develop any lesions during the 33 day follow-up period. Furthermore, viral genome copy numbers recovered in vaginal swabs from GEN-003-immunized animals were 100-fold lower than from controls. Recurrent shedding frequencies were reduced by 27%.
GEN-003 vaccination induced antigen-specific antibody responses at least as strong as those elicited by viral infection. Driven by gD-specific antibodies, HSV-2 neutralizing antibody titers followed the same pattern. After viral challenge, peripheral antigen-specific T cell responses in spleens of GEN-003-immunized animals were lower than those in controls, as measured by interferon-γ ELISpot. This might have resulted from increased antigen-specific T cell recruitment to the genital mucosa, or decreased exposure to the virus due to fewer recurrences. Additional methods to better understand mucosal mechanisms of protection are in development.
In summary, GEN-003 shows promise as a prophylactic vaccine candidate for genital herpes. Ongoing studies will determine if protection can be further enhanced by addition or exchange of antigens and/or adjuvants.
Vaccine-generated lung tissue-resident memory provides heterosubtypic protection to influenza infection
Authors: Zens Kyra D , Chen Jun Kui , Farber Donna L 
Published: January 2023

Abstract
Tissue-resident memory T cells (TRM) comprise a non-circulating subset which provides robust in situ protection upon infection. It is increasingly recognized that vaccination strategies promoting TRM may convey enhanced protection from disease. However, it is unclear whether currently available vaccines elicit TRM and whether this is dependent on vaccine formulation or administration route. In the context of influenza, we assessed lung TRM generation and subsequent protection following vaccination with either injectable inactivated influenza virus (IIV, Fluzone®) or intranasal live-attenuated influenza virus (LAIV, FluMist®) vaccines. We found that IIV induced primarily neutralizing antibodies while LAIV generated robust lung T cell responses. Importantly, LAIV, but not IIV, established persisting lung CD4 and CD8 TRM similar in localization and phenotype to that generated by influenza infection. Furthermore, protection experiments utilizing the immunomodulatory drug FTY720, which sequesters circulating T cells, including TEM and TCM, in secondary lymphoid tissues leaving lung TRM intact, demonstrated that only LAIV provided TRM-mediated protection to viral infection as evidenced by reduced weight loss and lung viral titers in this group following infection. Importantly, TRM generation and protection were dependent on both route and vaccine formulation. The ability of LAIV to generate lung TRM protective against non-vaccine viral strains may have important implications in providing protection in pandemic situations.
How Tolerance Shapes the Development of the HIV-1 Broadly Neutralizing Antibody 2F5
Authors: Finney Joel T , Yang Guang , Kuraoka Masayuki , Nojima Takuya , Verkoczy Laurent , Kitamura Daisuke , Haynes Barton F , Kelsoe Garnett H 
Published: January 2023

Abstract
An effective HIV-1 vaccine is likely to require induction of broadly neutralizing antibodies (bnAbs), which neutralize multiple HIV-1 strains. Generating HIV-1 bnAbs has been challenging due partly to a number of viral immune evasion mechanisms, including viral molecular mimicry of host in order to exploit immunological tolerance. Indeed, generation of neutralizing antibodies to the 2F5 epitope of HIV-1 gp41 is proscribed by immune tolerance. However, the specific mechanisms by which tolerance relieves the autoreactivity of B cells expressing the VH and VL regions (dKI) or the VH region (sKI) of the 2F5 bnAb are not well understood. To compare the reactivity and repertoire of 2F5 dKI and sKI B cells before and after immune tolerance checkpoints, we used a newly developed in vitro single B cell culture system that induces B cell expansion and differentiation into Ab-secreting plasmacytes. The reactivities of expanded B cell clones were determined by ELISA, while repertoire was determined by recovering the V(D)J rearrangements using RT-PCR and Sanger sequencing. In 2F5 dKI mice, the great majority of cells in the small-pre-B compartment express the knock in heavy- and light-chains, and maintain HIV-1 gp41-reactivity. However, 2F5 dKI B cells that survive tolerance checkpoints to become mature B cells have undergone extensive light-chain editing and are purged of gp41- and KYNU-reactivity. Importantly, the peripheral 2F5 dKI IgM−IgD+ (anergic) B cell subset retains a substantial fraction of HIV-1- and KYNU-reactive cells. In conclusion, our study provides mechanistic insights into how immunological tolerance impairs humoral responses to HIV-1, and suggests activation of anergic B cells as a potential target for HIV-1 vaccination.
Early prophylaxis impact for newborns at risk for HIV-1 transmission at birth: viral clearance and attenuation with neutralizing monoclonal antibodies in infant rhesus macaques
Authors: Hessell Ann Jones , Sacha Jonah B. , Kahl Christoph , Graham Barney , Mascola John R , Haigwood Nancy L 
Published: January 2023

Abstract
Our pre-clinical infant macaque model addresses translational implications for early intervention in mother-to-child transmission (MTCT). This model has shown that neutralizing monoclonal antibody (NmAb) therapy given 1 day after SHIVSF162P3 virus exposure intercepts the earliest stages of infection in lymphoid and mucosal tissues. Serial sacrifices revealed that within 2 weeks early viral foci in tissues were eradicated. All animals followed for up to 28 weeks were completely clear of virus infection with no viral rebound after CD8-depletion. Here, twelve 1-month-old rhesus macaques, negative for MHC class I alleles Mamu B*08, B*17 and A*01, were inoculated orally with SHIVSF162P3. Cocktails of NmAbs PGT121 and VRC07-523 were delivered subcutaneously at days 1, 4, 7, and 10 after SHIV exposure. Control groups did not receive NmAbs. Plasma, PBMC, and multiple tissues at necropsy were monitored for viral RNA and DNA. ICS was used to detect Gag and Vif T cell responses. We report that lowering NmAb dosing from 10 to 5 mg/kg given 24 h after virus exposure resulted in only one animal infected, with viremia persisting at about 1 log10 below controls. NmAb treatment at 10 mg/kg delayed for 48 h after oral SHIVSF162P3 exposure resulted in ablated virus infection in 3 of 6 macaques. CD8-depletion resulted in no evidence of virus rebound, indicating the absence of viral latency. In 2 macaques, NmAbs attenuated set-point viremia by 1 – 2 log10, compared to viremia in control animals. These results show that antibodies can have powerful and durable effects on viremia and disease in newborns, but timing and dose are critical. Moreover, post-exposure viral clearance from infected tissue marks a significant advance in pre-clinical platforms to prevent MTCT.
Vaccination route has an impact on level of protection of non-human primates from tuberculosis
Authors: Wong Eileen A , Agger Else Marie , Andersen Peter , Flynn JoAnne L 
Published: January 2023

Abstract
Tuberculosis (TB) continues to be a major global health problem, causing 1.5 million deaths in 2014. To curb TB, several TB vaccine candidates are currently in various stages of testing. We investigated the importance of vaccination route in immune response and protection against TB in a non-human primate (NHP) model. We vaccinated cynomolgus macaques with a protein subunit vaccine (H56) admixed with a novel adjuvant (CAF09) by three routes—intranasal, subcutaneous, and aerosol. Immunogenicity was measured by intracellular cytokine staining of blood and bronchoalveolar lavage samples during and after vaccination. NHPs were challenged with M. tuberculosis, and their immune recall responses assessed by blood and BAL samples. Lung inflammation was tracked during the course of infection by serial PET-CT scans, and extent of disease was assessed at necropsy. Frequencies of T cells producing pro-inflammatory cytokines increased in the blood and BAL following vaccinations in all vaccinated groups, but NHPs vaccinated by aerosol had an earlier recall of T cells to the airways after the third vaccination. After challenge, aerosol vaccinated NHPs had less inflammation by PET-CT compared to other vaccinated groups and unvaccinated controls. The aerosol cohort had similar gross pathology and total bacterial burden to controls at necropsy, but improved bacterial clearance and higher frequency of IFN-γ-producing T cells in the granulomas compared to controls. Although all vaccinated groups had TB disease, this study suggests that the route of vaccination affects the immune response and extent of disease, with the aerosol group having better outcomes. Further studies into the beneficial effects of aerosolizing TB vaccines are warranted.
Tight regulation of memory CD4 T cells limits immunopathology following chronic viral infection
Authors: Penaloza-MacMaster Pablo A , Barouch Dan H 
Published: January 2023

Abstract
CD4 T cells are critical for the induction of adaptive immune responses. Surprisingly, we published a report showing that memory CD4 T cells could also mediate lethal immunopathology following chronic lymphocytic choriomeningitis virus (LCMV) infection. To interrogate whether naive or memory CD4 T cells can similarly mediate immune dysregulation, we transferred equal numbers of naive or memory virus-specific CD4 T cells into naive mice, and infected these with chronic LCMV. We found that memory CD4 T cells exhausted substantially faster than naive CD4 T cells following chronic viral infection. This downregulation of adoptively transferred memory CD4 T cells was associated with improved adaptive immune responses and survival relative to naive CD4 T cells. Our findings demonstrate the importance of inducing multiple arms of the adaptive immune response and suggest that tight regulation of memory CD4 T cells is necessary for preventing immune disease during chronic viral infection.
The role of MAVS signaling in humoral immune responses to West Nile virus.
Authors: Young Lucy B , Giordano Daniela , Draves Kevin E. , Clark Edward A. 
Published: January 2023

Abstract
MAVS is a key regulator of innate immune responses to RNA viruses, such as West Nile virus (WNV). Previous work identified that MAVS is important for the development of adaptive immunity to WNV. MAVS KO mice are highly susceptible to infection with lethal WNV-Tx. MAVS KO mice produced significantly increased levels of WNV-specific IgM and IgG compared to WT mice, however, a corresponding increase in neutralizing antibodies (NAbs) was not observed. As MAVS KO mice succumb rapidly to WNV-Tx, it was not possible in our initial study to define the role of MAVS in the development of the germinal centers (GCs) as well as long-term plasma cell (PC) and memory B cell responses. Using the less pathogenic WNV-MAD, we examined the development of humoral responses in the absence of MAVS. MAVS KO mice survived infection with WNV-MAD, and as with WNV-Tx infection, produced increased levels of IgM and IgG but comparatively low levels of NAbs. This increase in serum Ab levels was maintained up to 9 weeks post-infection (pi). Moreover, the number of WNV-specific Ab secreting cells in the spleen and bone marrow were increased in MAVS KO mice compared to WT mice. Using flow cytometry we found that MAVS KO mice have a slight delay in expansion of GL7+ GC B cells, but by day 10 pi the number of GC B cells was about 10-fold higher than in WT mice. MAVS KO mice also showed a rapid expansion of IgM−CD138hi PCs that was significantly increased compared to WT mice. We are currently testing the hypothesis that these results are due to an intrinsic dysregulation of MAVS KO B cells rather than simply due to an increase in viral load. We are also examining both extrafollicular and GC responses to identify the basis for the rapid IgG antibody responses in MAVS KO mice. (Supported by NIH grant AI83019).
A tissue-specific phenotype for CD8+ T cells in atherosclerotic lesions: role of the arterial microenvironment
Authors: van Duijn Janine , Jiskoot Wim , Kuiper Johan , Slütter Bram 
Published: January 2023

Abstract
Atherosclerosis is a chronic, inflammatory disorder of the arterial wall that causes millions of deaths worldwide each year. CD8+ T lymphocytes may play an important role in atherogenesis, as they represent 29% of all lymphocytes in early human lesions, increasing up to 50% in advanced plaques, however their contribution to plaque initiation and progression remains unclear. Here we show that CD8+ T cells derived from enzymatically digested atherosclerotic lesions of ApoE−/− mice display a unique phenotype. Most notably, lesion derived CD8+ T cells produce less IFN-γ and TNF-α after PMA/Ionomycin stimulation compared to their counterparts in the spleen, circulation or other non-lymphoid organs. Importantly, cytokine production decreases as the lesion progresses to more advanced stages. This effect is not due to classical exhaustion of the T cells, as we found no difference in the expression of exhaustion markers such as PD-1. Nor is this effect due to an increase in regulatory FoxP3+CD8+ T cells. Together, our results suggest that the specific atherosclerotic environment in the vessel wall decreases CD8+ T cell functionality, which provides a new understanding of CD8+ T cells and their role in the pathogenesis of atherosclerosis.
The contribution of the HSCs niche to bone marrow failure in the IL-2 deficient model
Authors: Al-Kuhlani Mufadhal M , Gravano David M , Manilay Jennifer O , Hoyer Katrina K 
Published: January 2023

Abstract
Bone marrow failure (BMF) is a devastating disease where regeneration of peripheral blood lineages is compromised, resulting in anemia and mortality. In IL-2 KO BALB/c mice, the balance between early myeloid and lymphoid progenitor cell populations is skewed resulting in insufficient myelopoiesis with accumulation of CD8 T cells in the BM. It is widely accepted that crosstalk between cells of the bone microenvironment and hematopoietic cells affects each other’s behavior. However, it remains unknown how the BM niche in this mouse model contribute to the BMF. Multipotent stromal cells (MSCs), osteoblasts (OBs) and endothelial cells (ECs) are the three types of BM stromal cells (BMSCs) that may serve as the distinct functional niche to maintain HSCs. Repots have shown the interplay between CD8 T cells and MSCs cause a change in myelopoiesis and that MSCs promote the formation of Tregs. Our data also indicates that IL-2 KO CD8 T cells are important for the expansion of CD4 T cells in the bone. Since IL-2 KO bones are extremely brittle and CD4 T cells produce RANKL, the ligand needed for osteoclast (OC) differentiation, we evaluated the frequency of BMSCs and OCs. Our preliminary data shows clear differences in BMSCs profiles of IL-2 KO mice, namely elevated stromal cell numbers and a shift in Sca-1 expression in ECs and CD51 expression in OBs. Additionally, OC precursor frequency is elevated in the BM, indicating an increase in bone resorption. The ability of the different subsets of the BMSCs to support the differentiation of HSCs in vitro and the role of CD8 T cells in promoting these changes in vivo is being currently evaluated. Overall, our preliminary data suggests that the change in the bone microenvironment may contribute to the BMF observed in these mice.
Thymic expression of tissue-restricted self-antigens is a highly coordinated and evolutionary conserved process
Authors: Rattay Kristin , Meyer Hannah Verena , Brennecke Philip , Reyes Alejandro , Pinto Sheena , Brors Benedikt , Huber Wolfgang , Steinmetz Lars , Kyewski Bruno 
Published: January 2023

Abstract
Promiscuous gene expression (pGE) of tissue-restricted antigens (TRAs) in medullary thymic epithelial cells (mTECs) is essential for tolerance imposition in the thymus. PGE is characterized on the one hand by inclusion of a broad range of TRAs and on the other hand by its mosaic patterns, whereby each antigen is only expressed in 1–3% of mTECs at a given point in time. Yet, this mosaic pattern at the single cell level faithfully adds up to the full repertoire of self-antigens at the population level.
In order to analyze the regulatory mechanisms underlying this transcriptional heterogeneity among mTECs, we applied two complementing approaches, the isolation of minor mTEC subsets as defined by TRA-selected gene co-expression groups in conjunction with single cell mRNA sequencing.
Different TRA-selected mTEC subfractions, each expressing distinct sets of genes in a mutually overlapping fashion, mapped to distinct stages of mTEC development. These co-expression patterns were evolutionary conserved between mouse and human (Rattay et al., J. Autoimmunity 2015).
Applying an unbiased single cell mRNA sequencing approach, we extended these findings to the single cell level and showed that the mouse mTEC population essentially represents a composite of multiple co-expression groups (Brennecke et al., Nat. Immunology 2015).
These co-expression groups may represent only snapshots of a continuum of changing co-expression groups along the lifetime of an individual mTEC, as captured in the model of “sliding co-expression groups” (Pinto et al., PNAS 2013). Continuous genome scanning would potentially enlarge the overall diversity of self-antigens displayed by a single mTEC.
Inhibition of IL-23/IL-23 Receptor signaling ameliorates disease in lupus-prone mice by decreasing the generation of DN T cells
Authors: Dai Hong , Kyttaris Vasileios Christos 
Published: January 2023

Abstract
CD4−CD8−B220+ alpha beta (double negative T cells, DN) T cells are expanded in both SLE patients and lupus-prone mice. Based on our previous data (Zhang et al, J Immunol, 2009), we postulate that interleukin-23 is important for the generation of DN T cells. In the present study, we examined B220+ DN T cells in IL-23p19KO lpr mice and IL-23 receptor KO lpr mice by flow cytometric analysis; We also injected IL-23 into IL23p19KO lpr mice to reconstitute IL-23 expression. We observed that IL23p19KO or IL-23RKO lpr mice had reduced disease severity compared to control mice, accompanied by dramatically decreased CD4−CD8−B220+ alpha beta T cells in the peripheral lymph nodes and spleen. Using BrDU incorporation, we observed that there was enhanced proliferation of CD8+ T cells in the thymus of IL23p19KO or IL-23RKO lpr mice vs. controls. We also observed enhanced apoptosis of thymic DN T cells. IL-23 administration in IL23p19KO B6 lpr mice, but not in IL-23RKO B6 lpr mice, resulted in reconstitution of both CD8+T cells and DN T cell population in the periphery to similar levels as the wild type B6lpr mice. Thus, we conclude that deficiency of IL-23/IL-23R signal ameliorates disease in lupus-prone mice partly by decreasing the generation of DN T cells and normalizing CD8+ T cell levels.
Construction of vector pCAG-Z-eGFP-mIR for T cell specific expression of mouse IR
Authors: Al-Dieri Ali G , Morran Michael , McInerney Marcia F 
Published: January 2023

Abstract
Type 1 diabetes is a T cell mediated autoimmune disease. The insulin receptor (IR) is a chemotactic molecule capable of cell movement toward an insulin gradient. In NOD mice, T cells expressing IR can move into the islet causing insulitis and diabetes, whereas IR negative cells do neither. Our aim was to determine if a high density of IR expression is a viable mechanism to deliver T cells to the beta cells regardless of antigen specificity, in strains of mice that do not normally become diabetic. Therefore, we will use a Cre-Lox system with a strong CAG viral promoter and eGFP reporter to drive high IR expression on all T cells. To this end, a viable plasmid was constructed by sub cloning a mIR fragment into a lox vector with CRE recombinase restriction to ensure specific expression. Preliminary data shows successful transfection of this novel plasmid into a HEK293T CRE and NIH/3T3 cell line.
Listeria monocytogenes-encoded expression of a neuroantigen induces CD8 T-cell-mediated inhibition of autoimmune demyelinating disease
Authors: Itani Farah R. , Brate Ashley , Sinha Sushmita , Epping Lecia , Harty John T , Karandikar Nitin J 
Published: January 2023

Abstract
CD8 T-cells predominate in the central nervous system (CNS) lesions of multiple sclerosis (MS), an autoimmune demyelinating disease, yet their function remains controversial. We have previously shown that myelin-specific CD8 T-cells suppress experimental autoimmune encephalomyelitis (EAE), a murine model of MS. Here, we describe a novel system to probe the role of endogenously generated myelin specific-CD8 T-cell responses in CNS autoimmunity. A sequence encoding proteolipid protein peptide PLP178-191 was inserted into attenuated Listeria monocytogenes (LM), deleted for the actA and inlB genes. PLP178-191-specific CD8 T-cell responses were confirmed by infecting B6 mice with the LM-PLP strain and assessing peptide-induced IFN-γ production. We tested the effect of LMP-PLP infection on chronic (B6 mice) and relapsing-remitting (SJL mice) forms of EAE. In both systems, infection with LM-PLP did not result in disease induction. In contrast, LM-PLP infection resulted in significant amelioration of clinical disease in PLP178-191-immunized B6 and SJL mice, compared to LM-OVA-infected controls. LM-PLP infection also effectively suppressed relapses in SJL/J mice. Interestingly, LM-PLP-induced protection was not observed in CD8(−/−) B6 mice, indicating the importance of CD8 T-cells in mediating this effect. Our studies indicate that myelin-specific CD8 T-cell responses generated following LM-PLP175-194 infection are not encephalitogenic and attenuate CNS autoimmunity in multiple mouse models of EAE, further strengthening their role as regulators of disease and their potential as a novel immunotherapeutic strategy.