bluuebunny/crossref_metadata_2025_split huggingface dataset:
Co-Expression of Yersinia pestis F1- and V-Ags by the Same Salmonella Vaccine Protects Against Nasal Y. pestis Challenge (41.14)
Authors: Trunkle Theresa , Yang Xinghong , Bosio Catherine M. , Pascual David W. 
Published: January 2023

Abstract
To test whether the protective antigens (Ags) to Yersinia pestis can be simultaneously delivered, the Y. pestis cafI operon encoding the F1 Ag, and virulence Ag (V-Ag) were cloned into attenuated Salmonella vaccine vector. F1-Ag expression was controlled under its endogenous promoter from the cafI operon and amply expressed; a chimeric promoter consisting of ptetS and pphoP in the pV55 plasmid showed enhanced V-Ag expression. Immunofluorescence data showed that the single construct, Salmonella-(F1-V)Ags, could co-express both Ags. When orally tested, the Salmonella-(F1-V)Ags vaccine elicited elevated Ab titers to both F1- and V-Ags. Supportive CD4+ T cell responses showed that IFN-γ was elevated in response to F1-Ag restimulation and to a lesser degree, IL-4 and IL-13. V-Ag restimulation showed much less IFN-γ and IL-4 production. IL-10 responses were equivalent to both Ags. IgG1 and IgG2a subclass responses to both Ags were equivalent. Upon nasal challenge, immunized mice required immunity to both F1- and V-Ags because the mice vaccinated with Salmonella-(F1-V)Ags protected against 100 LD50 Y. pestis, but mice immunized with Salmonella constructs bearing a single Ag showed impaired efficacies. These results show that a single Salmonella vaccine can deliver both F1- and V-Ags to effect immune protection against Y. pestis.
This work is supported by AI-56286 and NIH 1U54 AI-06537.
Mucosal Immunity to Avian Influenza Following Ocular Immunization with Replication-Deficient Ad5-H5 in Chicken (41.15)
Authors: van GINKEL Frederik W. , Gulley Stephen L. , Toro Haroldo 
Published: January 2023

Abstract
Mucosal immunity plays an important role in the defense against viral pathogens. Limited information is available on the role of mucosal immunity in the chicken Harderian gland (HG) following ocular challenge. Although research has been performed on tears after ocular challenge or immunization of chickens, the mucosal immune response in the HG has not been well defined. To better understand mucosal immunity in HGs, we exposed chickens to a replication competent – free, recombinant adenovirus (Ad5) vaccine expressing the H5 gene of avian influenza (AI) virus. Expression of H5 in the HG after ocular administration of Ad5-H5 was demonstrated by immunohistochemistry. IgA and IgG H5- and Ad5-specific ELISPOT assays were developed to directly measure antibody-secreting B cells in HGs. The IgA and IgG spot-forming cells peaked on day 9 for Ad5 and day 11 for the H5 transgene. Ocular immunization induced Ad5- and H5-specific antibodies in the systemic compartment as measured by ELISA and HI assays, respectively. Increased levels of antibodies were also detected in tears. Western analyses demonstrated the presence of both monomeric and polymeric IgA in tears. RT-PCR analysis of HG derived mRNA confirmed the expression of the polymeric immunoglobulin receptor, confirming the importance of the HG as a mucosal effector site. Mucosal immunity was effectively induced by ocular application of the Ad5-expressed AI vaccine.
Intranasal immunization with Cry1Ac activates differentially lymphocytes in NALT as in NP. (41.16)
Authors: MORENO-FIERROS LETICIA , RODRIGUEZ-MONROY MARCO A. 
Published: January 2023

Abstract
Cry1Ac from B. thuringiensis is a potent mucosal immunogen and adjuvant. Via i.n. route Cry1Ac induces significant antibody responses and is able to improve vaccination against nasal invading pathogens, but their effects on nasal lymphocytes have not been determined. NALT and NP are considered as inductive and effector sites, respectively. We investigated the effect of i.n. immunization with Cry1Ac in the expression of CD69 and CD25 in lymphocytes of NALT and NP. Besides we analyzed effect in the proportion of T cells producing IL2, IL4, IL5, IL10, IFNγ and TNFα. Immunization with Cry1Ac increase considerably the expression of activation markers in both nasal tissues. But as expected for a mucosal effector site, NP contained major proportions of B220, T CD4 and T CD8 cells expressing CD25 and CD69 in comparison to NALT, Likewise, Cry1Ac increased the proportion of T cells spontaneously producing IL2, IFNγ, and IL4, and the effect was higher in NP than in NALT. Present study provides evidence that intranasal immunization with Cry1Ac activates differentially lymphocytes in NALT as in NP. These data support that Cry1Ac is a potent immunogen and a valuable study tool to the induction of nasal immune responses. The new information presented, is important, as it is necessary to have a thorough understanding of the unique molecular and cellular properties of the nasal immune system for the development of successful nasal vaccines
CONACyT 43102 and UNAM DGAPA PAPIIT IN213903 grants
Development of needle-free vaccine: rice-based oral vaccine induced protective immunity against cholera toxin (41.4)
Authors: Nochi Tomonori , Yuki Yoshikazu , Takagi Hidenori , Yang Lijun , Masumura Takehiro , Mejima Mio , Nakanishi Ushio , Matsumura Akiko , Hiroi Takachika , Morita Shigeto , Tanaka Kunisuke , Takaiwa Fumio , Kiyono Hiroshi 
Published: January 2023

Abstract
Capable of inducing antigen-specific immune responses in both systemic and mucosal compartments without the use of syringe and needle, mucosal vaccination is considered ideal for the global control of infectious diseases. In this study, we developed a rice-based oral vaccine expressing cholera toxin B subunit (CTB). An average of 30 μg of CTB per seed was stored in the protein body, a rice storage organelle. When orally fed, rice seeds expressing CTB were taken up by the M cells covering the Peyer′s patches (PPs), inducing toxin-specific serum IgG and mucosal IgA antibodies with neutralizing activity. When expressed in rice, CTB was protected from pepsin digestion in vitro. Rice-expressed CTB also remained stable and thus maintained immunogenicity at room temperature for more than 1.5 years, meaning that antigen-specific mucosal immune responses were induced at much lower doses than were necessary with purified recombinant CTB. Because they would require neither needles nor refrigeration (cold-chain management), these rice-based oral vaccines offer a highly practical as well as cost-effective strategy for orally vaccinating large populations against mucosal infections, including those which may result from an act of bioterrorism.
Th2 polarizing helminth infection alters the immune response to an oral Salmonella-OVA vaccine (41.5)
Authors: Iweala Onyinye I. , Smith Donald W. , Urban Joseph F. , Nagler Cathryn R. 
Published: January 2023

Abstract
We have previously shown that enteric helminth infection acts as a mucosal adjuvant, priming for a Th2 biased response to soluble ovalbumin (OVA), a tolerogenic form of dietary antigen. An immunogenic response to orally administered OVA can be elicited by administration of OVA in the context of an oral vaccine. To examine whether chronic enteric infection can also influence the efficacy of oral vaccination we examined the response to a novel OVA-expressing oral Salmonella vaccine (Salmonella-OVA; Smith et al submitted) in both infected and non-infected mice. This live attenuated vaccine induces a Th1 biased mucosal and systemic OVA specific response. Infection of C57Bl/6 mice with the helminthic parasite Heligmosomoides polygyrus delayed and reduced the Th1 dependent serum OVA specific IgG2c response induced by two doses of oral vaccine. By contrast, helminth infection enhanced the Th2 dependent serum OVA specific IgG1 response. Helminth infection also altered mucosal and systemic OVA specific cytokine and chemokine responses. These data suggest that concomitant enteric helminth infection can significantly alter the immune response to oral vaccination. Since chronic enteric parasitic infections are endemic in much of the developing world, our observations have important implications for the testing and development of mucosal vaccines.
This work was supported by NIH grants DK 55678 and DK43551 and predoctoral training fellowships 5F31AI054229-05 and T32 A107529.
Small and large intestinal intraepithelial T lymphocytes show distinct dependency on sphingosine 1-phosphate (42.11)
Authors: Kunisawa Jun , Kurashima Yosuke , Higuchi Morio , Gohda Masashi , Ishikawa Izumi , Ogahara Ikuko , Kim Namju , Shimizu Miki , Kiyono Hiroshi 
Published: January 2023

Abstract
It is known that the composition of intraepithelial T lymphocyte (IEL) differs between small and large intestines, but the mechanism underlying that difference remains obscure. Here, we show that sphingosine 1-phosphate (S1P) plays a key role in regulating intestinal IEL trafficking into the small and large intestines. High levels of type 1 S1P receptor (S1P1) expression was noted on naïve IELs expressing CD4 or CD8αβ, which leads to their preferential migration into the large intestine. In contrast, recent thymic emigrants (RTEs), double-positive thymocytes, and double-negative thymic T cell-committed precursors use S1P-independent trafficking pathway into the intestine. The former two populations exclusively migrate into the small intestine, while the latter double-negative thymic T cell-committed precursors migrate into both the small and large intestines. Hence, down-regulation of S1P1 expression inhibited naïve IEL migration into the intestines but did not affect the migration of thymic IEL precursors. These data are the first to demonstrate that a lipid-mediated system determines whether IELs migrate to the small or large intestine.
Placental cytotrophoblast HLA-G5 is synthesized as a β2m-free heavy chain homodimer (42.13)
Authors: Morales Pedro J , Pace Judith L , Platt Jeralyn S , Hunt Joan S 
Published: January 2023

Abstract
Placental villous cytotrophoblast cells (vCTB) contain an unusual member of the HLA class I gene family, HLA-G5. Recently we showed that HEK293-derived recombinant HLA-G5 protein is predominantly composed of H:H homodimers. Here, we examined the HLA-G5 produced in primary vCTB. Determining the structure is critical because recombinant ectodomains of the inhibitory leukocyte Ig-like LILRB1 (ILT2) and LILRB2 (ILT4) receptors bind HLA-G dimers with higher affinity than monomers. Analysis of vCTB cell extracts by immunoblotting demonstrated that vCTB cell HLA-G5 H chain proteins are primarily disulfide bonded dimers. The H chains were not associated with β2m L chains; although β2m mRNA was identified by RT-PCR, immunoblots failed to detect β2m protein even when additional β2m mRNA was introduced. Furthermore, sequencing failed to reveal any abnormalities in the translational start codon of either endogenous β2m mRNA or introduced β2m mRNA, and EGF-induced syncytialization did not promote β2m in vCTB cells. The inability of vCTB cells to translate β2m messages in vitro was reflected in failure to detect β2m in vCTB cells in situ. Thus, vCTB cells may synthesize a disulfide-bonded, β2m-free homodimeric form of HLA-G5 due to an inability to synthesize β2m protein. This molecule could comprise a particularly effective tolerogenic molecule at the maternal-fetal interface.
Late Pregnancy Serum Factors Suppress T Cell Activation in Experimental Autoimmune Encephalomyelitis (EAE) (42.14)
Authors: Gatson NaTosha N , McClain Melanie , Gienapp Ingrid E , Powell Nicole , Shawler Todd M , Kithcart Aaron P , Whitacre Caroline C 
Published: January 2023

Abstract
Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the CNS thought to be due to an autoimmune response directed against myelin antigens. EAE is a commonly used animal model for MS and shares clinical, histopathologic and immunologic similarities with MS. Profound suppression of the MS and EAE relapse rate is observed during late pregnancy and is followed by a marked increase in disease severity postpartum. We have recently observed that histopathologic progression of EAE is diminished during late pregnancy and is exacerbated post partum. During EAE and late pregnancy, mice were found to have less mononuclear CNS infiltrates, demyelination and axon severing than virgin controls. The purpose of this study was to explore the hypothesis that decreased autoimmune disease during pregnancy is due to immunosuppressive factors present in the serum during late pregnancy. Female SJL/J mice were immunized for EAE using myelin proteolipid protein (PLP) 139–151. T cell proliferation assays were carried out using lymph node (LN) cells stimulated with PLP or anti-CD3 in the presence of 0–3% late pregnancy or virgin mouse sera. LN cells from age and gender matched naïve mice served as controls. Three percent late pregnancy serum suppressed both anti-CD3 and PLP stimulated T cell proliferation as compared to cells cultured with virgin mouse sera. These results demonstrate the broad suppressive potential of late pregnancy serum on T cell activation and may be responsible for amelioration of clinical signs of EAE during late pregnancy.
(Supported by NIH grants NS48316 and T32AI055411).
Immunomodulatory actions of Treg PPAR γ in the gut mucosa (42.3)
Authors: Hontecillas Raquel , Bassaganya-Riera Josep 
Published: January 2023

Abstract
Peroxisome proliferator-activated receptor (PPAR) γ activation has been implicated in the prevention of immunoinflammatory disorders, however the mechanisms of immune modulation by endogenous PPAR γ agonists remain unclear. We have used PPAR γ-deficient CD4+ T cells obtained from tissue-specific PPAR γ null mice (i.e., PPAR γ fl/fl; MMTV-Cre+) to investigate the role of endogenous PPAR γ on regulatory CD4+ T cell (Treg) function. Overall, we show that the transfer of purified PPAR γ null CD4+ T cells into SCID recipients results in enteric disease. To test the assertion that the deficiency of PPAR γ in Treg impairs their ability to prevent effector T cell-induced colitis, we performed co-transfer studies using PPAR γ null and PPAR γ-expressing CD4+ T cells. These studies demonstrate that PPAR γ-expressing, but not PPAR ? null Treg, prevent colitis induced by transfer of naïve CD4+ T cells into SCID recipients. In line with these findings, the production of IFN-γ by spleen and mesenteric lymph node-derived CD4+ T cells was down-regulated following transfer of PPAR γ-expressing, but not PPAR γ null, Treg. Furthermore, the loss of PPAR γ limited the expansion of Foxp3+ cells systemically and their accumulation in the colonic mucosa. In conclusion, our data suggest that endogenous PPAR γ activation represents a Treg intrinsic mechanism of downregulation of effector CD4+ T cell function and prevention of intestinal inflammation.
Role of NKT cells in the placenta and uterus of pregnant mice infected by Chlamydia trachomatis (42.5)
Authors: Kanellopoulos-Langevin Colette , Delarbre Christiane , Habbeddine Mohamed , Verbeke Philippe , Moutier René , Ojcius David 
Published: January 2023

Abstract
In order to study the role of maternal NKT cells against a Chlamydia infection during pregnancy, we have compared CD1d-deficient, C57Bl/6 (B6) mice to wild type controls, in the presence or absence of a Chlamydia infection. We have examined the expression of several molecules known to be important for NKT cell function, in the uterus and placenta. We focused our study on NKG2A, NKG2C and NKG2D receptors, their ligands Qa-1 and Rae-1 and their co-receptors CD94, DAP10 and DAP12. We also studied perforin, granzyme B, osteopontin and cytokines such as TNF alpha, interferon gamma and IL1 beta, Our results suggest that CD1d-restricted NKT cells do not play a major role in the control of infection, as the intensity and duration of infection were comparable in CD1d-deficient and wild type mice. However, we found that Chlamydia infection caused an increase of invariant NKT (iNKT) cells in the placenta and uterus of B6 mice, measured by the expression of their characteristic TCR Valpha14-Jalpha18 invariant chain. Correlatively, Chlamydia infection in CD1d-deficient mice lead to organ-specific modulations in the expression of several NKT cell markers, which differed from those observed in B6 mice.
Supported by NIH grant number 5 R21 A1060725-02, CNRS and Universities Paris 6 and 7.
CXCL9 and CXCL10 Expression are Critical in Resistance to Genital Herpes Simplex Virus Type 2 (HSV-2) Infection (43.15)
Authors: Thapa Manoj , Carr Daniel 
Published: January 2023

Abstract
The relevance of CXCL9 and CXCL10 expression in protection against genital herpes simplex virus type 2 (HSV-2) infection was investigated using mice deficient in CXCL9 (CXCL9−/−) or CXCL10 (CXCL10−/−) along with C57BL/6 wild type (WT) mice. Progesterone-sensitized mice infected with 2,000 pfu HSV-2 intravaginally were exsanguinated at times post infection (p.i.), and the vaginal tissue (VG), spinal cord (SC), and brain stem (BS) were removed and assayed for virus titer, leukocytes infiltration, and cytokine/chemokine content. CXCL10−/−, but not CXCL9−/− or WT mice were found to possess HSV-2 in the SC at day 3 following infection. By day 7 p.i., CXCL9−/− and CXCL10−/− mice harbored significantly more virus in the SC and the BS compared to WT mice. The increase in virus titer in the BS and SC of the chemokine deficient mice correlated with a decrease in NK cell mobilization to the nervous system at day 5 p.i. and CD8+ T cells mobilization at day 7 p.i. In addition, there was a noticeable increase in vaginal inflammation (measured by cytokine/chemokine levels and leukocyte infiltration) in the CXCL10−/− mice compared to the WT or CXCL9−/− mice. Antibody-mediated depletion of NK cells resulted in an increase in HSV-2 levels in the vagina, SC, and BS of wild type mice suggesting these cells contribute to resistance to infection.
This work supported by AI053108.
The Effects of Listeria monocytogenes Infection on Dendritic Cell Subsets (43.16)
Authors: Mitchell Latoya M. , Grayson Jason , Hiltbold Elizabeth M. 
Published: January 2023

Abstract
Dendritic cells (DC) are antigen presenting cells that link innate and adaptive immunity and have been shown to be a vital component for clearance and protection against Listeria monocytogenes (Lm) infection. Because of the role of DC in immunity against Lm and the varied functional capacities of DC subsets, our goal is to determine the impact of Lm infection on CD8α+/CD24+, myeloid, and plasmacytoid DC subsets found in Flt3L bone-marrow derived DC (BMDC) and mouse spleen.
Previously, we found that Flt3L BMDC undergo maturation upon infection with wild-type Lm as well as a Hly− mutant Lm that cannot enter the cytoplasm. The response in Flt3L BMDC to Hly− Lm is also dependent on signaling through the Toll-like receptor adaptor, MyD88. These data are in contrast to GM-CSF BMDC which only mature in response to cytoplasmic Lm. In vivo studies showed that the peak of costimulatory molecule upregulation following i.v. infection with wild-type Lm is 24 hours. Splenic DC, in contrast to Flt3L BMDC, did not increase surface costimulatory molecules in response to Hly− Lm. The frequency of splenic DC subsets following Lm infection and their ability to present antigen to T cells was also studied. In summary, our goal is to understand the capacity of different DC subpopulations to handle Lm infection and in future studies determine the role of each DC subset in establishing protective immunity. (NIH AI057770)
Lytic granule components are required for the maintenance of HSV-1 neuronal latency (43.35)
Authors: Knickelbein Jared E , Khanna Kamal M , Hendricks Robert L 
Published: January 2023

Abstract
Using a murine model of HSV-1 corneal infection, our data suggest that secretion of CD8 T cell lytic granules (LGs), in addition to IFN-γ, is required for the maintenance of viral latency within trigeminal ganglia (TG). First, HSV-specific CD8 T cells (HSV-CD8), which recognize an immunodominant HSV-1 glycoprotein B (gB498–505) epitope, surround neurons within latently infected TG and contain functional LGs, with GrB polarizing to the contact site with these neurons. HSV-CD8 use GrB and IFN-γ to block HSV-1 reactivation from latency in a non-redundant manner in ex vivo TG cultures, possibly due to a lack of IFN-γ receptors on some latently infected neurons. Like WT mice, Pfn and GrB KO mice establish latency in the TG by 8 days post-infection (dpi) with a similar viral genome copy number, but by 14 dpi, they exhibit a transient rise in viral genome copy number not seen in WT mice suggestive of HSV-1 reactivation from latency. A similar transient rise in copy number is also seen in WT mice depleted of CD8 T cells. Further, neither endogenous CD8 T cells within Pfn KO nor exogenous HSV-CD8 from GrB KO latently infected TG maintain latency as efficiently as CD8 T cells from WT TG in ex vivo TG cultures. Based on the current lack of evidence that CD8 T cell protection leads to neuronal death, we hypothesize a non-lethal LG-mediated block in HSV-1 reactivation from latency and are currently investigating possible mechanisms of protection.
NKG2D Regulates Viral Load in the CNS During Acute Picornavirus Infection (43.37)
Authors: Deb Chandra , Howe Charles 
Published: January 2023

Abstract
Neurovirulent picornaviral infections are a significant health risk in developing nations. Unfortunately, the mechanisms of picornaviral clearance from the central nervous system (CNS) are less understood and therapeutic approaches are limited. We have established a mouse model of neurovirulent picornavirus infection using the Theiler’s murine encephalomyelitis virus (TMEV) which replicates during the first 12 days postinfection (dpi) primarily in the hippocampus but is cleared from the brain by 21 dpi. Intracerebral infection of TMEV triggers the early infiltration of natural killer cells (NKCs) followed by CD8+ T cells (CTLs) into the brain. We observed that both brain-infiltrating NKCs and CTLs express the NKG2D activating receptor on their surface and 50% of the the CTLs specific to the VP2121–130 viral capsid peptide also expressed NKG2D. Concomitantly, brains of infected mice expressed NKG2D ligands such as RAE-1, H-60 and MULT-1 and microglia were the cellular locus of these ligands. Importantly, systemic treatment with the NKG2D function blocking antibody CX5 resulted in a significant increase in viral load in the brain. These results suggest that activated and infected microglia stimulate NKCs and CTLs via NKG2D in a manner that restricts viral replication. NKG2D may thus serve as a potential therapeutic target to enhance viral clearance responses during CNS infection.
This work was supported by a grant from the National MS Society and by Donald and Frances Herdrich.