bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inflammatory chemokine CCL3 suppresses preferential tumor-associated accumulation of B cells and CD4 T cells in tumor-draining lymph nodes (P2017)
Authors: Allen Frederick , Bobanga Iuliana , Barkauskas Deborah , Myers Jay , Tsao Matt , Huang Alex 
Published: January 2023

Abstract
We have previously demonstrated that CCL3 (MIP-1α) and CCL4 (MIP-1β) were important in orchestrating cellular contacts in vaccinated lymph nodes (LNs) and can enhance memory T cell generation (Castellino et al). We hypothesize that incorporating CCL3 and CCL4 into tumor vaccines can enhance anti-tumor immunity. Using colon tumor model CT26, we generated tumors that secret CCL3, CCL4 or both, and inoculate them live into the footpad of naïve recipients. CCL3-secreting tumors failed to grow in immunocompetent mice, and enhance antitumor immunity when used as a vaccine. Flow cytometric analysis of draining popliteal LNs 3 days after live cell injection revealed that while the total LN cell number increased by 2.03 fold after wild type (WT) tumor challenge relative to contralateral LN (conLN), CCL3-secreting tumor injection failed to do so despite the presence of tumor cells in the LNs. CD4 cells in the WT draining LN were 1.58 fold higher compared to conLN, but remained unchanged in CCL3-secreting tumors. Striking differences were seen in B cells where WT tumor resulted in a 7.04-fold increased, but CCL3-secreting tumors only caused a modest 2.04-fold change. Dynamic intravital 2-photon microscopy and tissue histology studies further revealed that tumors occupy the B cell follicle during early metastasis, and exogenous CCL3 gradient may potentially alter lymphocyte homeostasis in tumor draining LN, resulting in immune activation rather than tolerance against CT26.
Pan-Shigella Surface Protein antigen 1 as potential cross-protective antigen for Shigella vaccine development. (P4286)
Authors: Kim Jae-Ouk , Kim Su Hee , Rho Semi , Kim Heejoo , Dey Ayan , Yang Jae-Seung , Nandy Ranjan , Czerkinsky Cecil , Kim Dong Wook 
Published: January 2023

Abstract
Shigellosis is a disease characterized by the destruction of the colonic epithelium in humans and one of the most severe diarrheal diseases, especially in children under 5 years of age in developing countries. We have identified several protein antigens that are shared by different shigella serotypes and species. Among these, one antigen, PSSP-1, induces cross-protection against experimental shigellosis induced by different species and serotypes. PSSP-1-induced protection requires (i) a mucosal route of administration; and (ii) the co-administration of a mucosal adjuvant (cholera toxin, mutant thermolabile enterotoxin (dmLT). When PSSP-1was formulated with dmLT, an adjuvant that has undergone human clinical testing, it induced cross-protection against S. flexneri 2a, 3a, 5a, 6, S. boydii and importantly also S. dysenteriae 1. While intradermally administered PSSP-1 adjuvanted with dmLT induced strong serum antibody responses, it failed to induce protection in the mouse lung pneumonia model (consisting of administering Shigella intranasally). In contrast, intranasal PSSP-1 + dmLT elicited efficient local (lung) and peripheral (spleen) antibody responses and regional and systemic IL-17 and IFNγ production. Interestingly, patients with recent onset shigellosis showed variable but significant antibody responses to most conserved Shigella protein antigens but not to PSSP-1. Although the above results are promising, these will have to be validated in humans.
Fc receptor-targeted vaccines: an adjuvant-independent approach for mucosal vaccination against intracellular and extracellular mucosal pathogens (P4290)
Authors: Gosselin Edmund , Bitsaktsis Constantine , Iglesias Bibiana , Pham Giang , Drake James , Snapper Clifford , Franz Brian 
Published: January 2023

Abstract
We previously demonstrated that immunization with inactivated Francisella tularensis, a Category A intracellular mucosal pathogen, combined with anti-F. tularensis mAb, enhances protection against F. tularensis challenge. We thus sought to determine the mechanism(s) involved, and if this strategy is compatible with recombinant vaccine technology. We found that inactivated F. tularensis combined with F. tularensis-specific mAb extends immunogen presentation to T cells via FcγR-mediated Ag persistence, enhances transport of immunogen to the NALT, increases recruitment of DCs to the lung, and increases the number of CD4 and IFN-γ producing effector memory T cells. Utilizing an extracellular infectious disease model (S. pneumoniae), we also demonstrate this strategy can be applied as a recombinant fusion protein composed of the immunogen PspA targeted to human FcγRI (constitutively expressed on DCs and macrophages). Furthermore, traditional adjuvant is not required to enhance protection in either vaccine model. In summary, for the first time, we utilize an established mucosal vaccine protection model to identify mechanisms involved in FcR-enhanced mucosal immunity. We also demonstrate this strategy is effective against both intracellular and extracellular mucosal pathogens and is compatible with the use of recombinant vaccine technology. These studies thus establish the broad applicability of FcR-targeted immunogens as an effective adjuvant-independent mucosal vaccine platform.
Efficacy of a prime/boost regimen including BCG and recombinant protein in an HN878 challenge of C57BL/6 mice (P4296)
Authors: Schaeffer Jacqueline , Laddy Dominick , Wachholder Robert , Anantha Ravi , Imam Zakari , Morris Sheldon , Derrick Steven , Hokey David 
Published: January 2023

Abstract
Tuberculosis remains a major concern despite the widespread use of BCG vaccine. Since BCG-induced protection against adult pulmonary TB has been highly variable, vaccines that confer better, more complete protection are needed and recombinant proteins in the presence of adjuvant are currently being evaluated in this context. C57BL/6 mice were used in an immunogenicity and aerosol challenge study to evaluate the efficacy of a heterologous prime-boost vaccination regimen. Adult females aged 6-8 weeks old were vaccinated with either BCG SSI alone, or in conjunction with a recombinant fusion protein expressing Ag85B, Ag85A and Rv3407 with a poly(I:C)-based TLR3 adjuvant. Prior to challenge, a subset of mice were used to evaluate immune responses by IFN-γ ELISA and flow cytometry. This combination elicited a strong Th1 response as evidenced by high levels of IFN-γ, IL-2 and TNF expression in CD4+ T-cells from splenocytes. Mice were challenged with 50-100 CFU of aerosolized M.tuberculosis strain HN878. Spleens and lungs were used to determine viable bacterial load (CFU) and lungs fixed for routine histopathology. We observed a statistically significant reduction in lung CFU at 12 weeks post infection in the prime-boost vaccine group compared to BCG alone. The use of a recombinant protein plus adjuvant as a boost to the widely used BCG SSI vaccine was shown to be immunogenic and to elicit an increased level of protection over BCG alone in the lungs of adult C57BL/6 mice.
In vivo targeting of human DC-SIGN drastically enhances CD8+ T cell-mediated protective immunity (P4298)
Authors: Berod Luciana , Hesse Christina , Ginter Wiebke , van Kooyk Yvette , Sparwasser Tim 
Published: January 2023

Abstract
Vaccination is one of the oldest and still most effective methods to prevent infectious diseases. Yet, eradication of intracellular pathogens and other diseases like cancer, requiring strong cytotoxic (CTL) T cell responses, remains a medical challenge. In mice, targeting antigens to dendritic cells (DCs) via specific antibodies against C-type lectin receptors represents a successful approach to induce strong cytotoxic immune responses if used in combination with adjuvants. However, a major drawback to use this strategy in the clinic is the lack of analogous targets in human DCs. DC-SIGN (DC specific ICAM3 grabbing non integrin/CD209) is a C-type lectin receptor with strong endocytic capacity and a highly restricted expression on human immature DCs. Therefore, DC-SIGN represents an ideal candidate for DC targeting. Using transgenic mice that express the human DC-SIGN receptor under the control of the murine CD11c promoter (hSIGN mice), we explored the efficacy of anti-DC- SIGN antibodies to target antigens to DCs and to induce protective immune responses against intracellular infectious agents like Listeria monocytogenes or Mycobacterium tuberculosis in vivo. We show that anti-DC-SIGN conjugated antibodies can induce strong and persistent antigen-specific CD4+ and CD8+ T cell responses and propose DC-SIGN as a promising candidate for novel vaccine strategies based on DC targeting against intracellular pathogens.
The role of Th17+ T cells and neutrophils in mucosal immunity against Staphylococcus aureus mastitis. (P4256)
Authors: Kanevsky-Mullarky Isis , Lehtimäki Mari , Wark Wendy , Mwangi Waithaka 
Published: January 2023

Abstract
Staphylococcus aureus is a common cause of mastitis, a disease estimated to cost the dairy industry ~$2 billion/yr. Public concern over the use of antibiotics in agricultural animals necessitates the identification of new mechanisms for controlling this pathogen. Our goal is to define the role of immune activating DC in the development of protective S. aureus-specific T cell memory and use this knowledge for identification of antigens for vaccine development. Our findings indicate IL-17 generated by PBMC in response to S. aureus loaded monocyte-derived DC that is greater in memory as compared to naïve animals. Lymphocytes stimulated with both live and gamma irradiated antigens presented by monocyte-derived DC released active Granzyme B. Cells from previously infected animals had higher Granzyme B response than animals with no previous infection. In animals with no previous clinical infections, irradiated S. aureus antigen induced lower response than live S. aureus. Furthermore, bovine neutrophils generate greater levels of cytokines when stimulated with supernatants from S. aureus induced T cells. This data indicates that previous infection can heighten the ability of cytotoxic T cells to produce Granzyme B and increase neutrophil function. The ability to increase presence of protective memory immune cells and innate immune cell function in the mammary gland will provide new vaccine targets for S. aureus.
Flagellin encoded in gene-based vector vaccines is a route-dependent immune adjuvant (P4302)
Authors: Rady Hamada , Huang Weitao , Ramsay Alistair 
Published: January 2023

Abstract
TLRs play an important role in the initiation of immune responses. Their ligands may, therefore, have adjuvant properties in vaccine settings. Flagellin (TLR5L) promotes strong antigen-specific humoral responses, however its capacity to enhance specific T cell responses is controversial. Here, we evaluate adjuvant properties of flagellin in gene-based vaccination strategies. Intramuscular (IM) immunization with DNA or Ad vaccine vectors encoding Ag85B of M. tuberculosis (Mtb) and flagellin enhanced Ag85B-specific CD4+ and CD8+ T cell responses over vectors encoding Ag85B alone. In contrast, intranasal (IN) co-immunization with Ad encoding flagellin led to decreased numbers of mucosal Ag85B-specific CD4+ and CD8+ T cells. Following DNA/Ad prime-boosting, mice co-primed with DNA-Ag85B-flagellin IM and then boosted with Ad-Ag85B IM or IN, had significantly enhanced Ag85B-specific CD4+ (IM and IN boosting) and CD8+ (IM boosting) T cell responses without obvious morbidity. Moreover, co-priming with flagellin enhanced protection against pathogenic aerosolized Mtb. Our data indicate that the capacity of flagellin to enhance Ag85B-specific cellular immune responses is route dependent, either when encoded in DNA or Ad vaccines, or following DNA/Ad prime-boosting. Inclusion of flagellin at the priming phase of DNA/Ad prime-boost vaccination has strong adjuvant activity enhancing protective immune responses to co-expressed vaccine antigen.
Stability of vaccine induced Th2 cells following intranasal infection with group A streptococcus (P4258)
Authors: Dileepan Thamotharampillai , Scully Ingrid , Anderson Annaliesa , Jansen Kathrin , Cleary Paul 
Published: January 2023

Abstract
Group A streptococcus (GAS) naturally colonizes the human pharynx and can cause pharyngitis. We have found that intranasal administration (i.n.) of GAS elicits a Th17 response in mice; in contrast, intramuscular (i.m.) injection of recombinant GAS protein vaccines elicits a Th2 response. The influence of GAS infection on the vaccination-induced Th2 population was assessed using IL-4 reporter mice (4get and KN2) immunized with a GAS protein vaccine plus alum. Vaccination caused expansion of IL-4+ T helper cells which did not express IL-17A. Purified GFP+IL-4+ Thy1.1+ CD4+ T cells from vaccinated 4get mice were adoptively transferred to Thy1.2+ congenic recipients to evaluate their response to subsequent GAS infections. Recipient mice were infected i.n. with GAS and cytokine profiles of transferred cells were determined. In 5 of 6 recipient mice, GFP+Thy1.1+ cells retained their original Th2 phenotype and did not express IL-17A. In the sixth mouse, 8.6% of Thy1.1+ cells expressed IL-17A after exposure to GAS. As expected, endogenous Thy1.2+ cells from infected recipients expressed IL-17A. Thus, im vaccination primarily induced Th2 cells, and most cells were terminally differentiated and unable to develop the capacity to express IL-17A upon subsequent mucosal exposure to GAS. Thus vaccination with recombinant GAS proteins is unlikely to create a reservoir of GAS-specific T cells that can become polarized toward a Th17 response upon GAS infection.
Systemic immunization with a recombinant Pneumocystis protein vaccine candidate induces systemic and mucosal immune responses in non-human primates (P4260)
Authors: Kling Heather , Burton Andrew , Mcmanus Alexandria , Guyach Siobhan , Tarantelli Rebecca , Norris Karen 
Published: January 2023

Abstract
Despite widespread antiretroviral therapy, pulmonary disease remains a leading cause of morbidity and mortality in HIV+ patients, likely resulting from increased colonization rates by Pneumocystis (Pc) jirovecii, which causes pneumonia and is associated with chronic obstructive pulmonary disease (COPD). We have previously shown that early detection of antibodies to Pc kexin-like protease (KEX1) in the bronchoalveolar lavage fluid (BALF) correlates with protection from Pc colonization and subsequent COPD development in a macaque model of HIV. Here, we test the capacity of a systemic recombinant KEX1 vaccine to induce detectable KEX1-antibody responses in the lungs. Rhesus macaques (n=6) were intramuscularly (i.m.) immunized with 100ug of truncated, recombinant KEX1 protein + aluminum hydroxide (Alum), rested, then boosted with 50ug of KEX1 + Alum. Mock-immunized macaques (n=6) were injected (i.m.) with proteins from empty E.coli expression vector (pET28b) + Alum. Following boost inoculation, all macaques had plasma KEX1-IgG RET &gt;250,000 (range=256,000-2,000,000) and BALF KEX1-IgG RET &gt;1800 (range=1900-31,800). Five of 6 macaques had detectable BALF KEX1-IgA (range=100-550). Thus, systemic immunization with recombinant protein and Alum results in significant boosting of both circulating and mucosal KEX1 antibody titers. These data support KEX1 as a potent vaccine candidate against Pneumocystis infection.
Are antibodies necessary for a Chlamydia T cell vaccine? (P4278)
Authors: Karunakaran Karuna , Yu Hong , Jiang Xiaozhou , Li Yuling , Shen Caixia , Brunham Robert 
Published: January 2023

Abstract
The murine Chlamydia muridarum model demonstrates that CD4 T cells are necessary and sufficient to resolve primary infection and that either CD4 T cells or antibody is necessary and sufficient to resolve reinfection. It remains uncertain whether a Chlamydia vaccine should accelerate clearence or prevent reinfection. Using immunoproteomics we have identified five Chlamydia outer membrane proteins (PmpG, PmpE, PmpF, PmpH and MOMP) that when individually used as protein vaccine accelerated clearance of Chlamydia infection. All 5 proteins are located in the chlamydial outer membrane complex (COMC). We tested PmpG and MOMP (major outer membrane protein that constitute more than 60% of COMC) as recombinant proteins or in native conformation as part of COMC preparations for their efficacy in accelerating clearance, providing significant protection and reducing tubal pathology. Our results show that COMC, PmpG and MOMP formulated with a Th1 polarizing adjuvant accelerated clearance but that only COMC exhibited near complete protection to challenge infection and reduced genital tract pathology. High titres of Chlamydia neutralizing antibodies were only detected following COMC immunization and not following PmpG or MOMP immunization. The superior efficacy of COMC as Chlamydia vaccine may be due to native conformation of outer membrane proteins that induce functional antibodies as well as CD4 T cell responses. These results suggest COMC may be a promising Chlamydia vaccine vehicle.
High plasma TNF-α levels and TNF-α/IL-10 ratios are associated with TNF2 allele in severe P. falciparum malaria patients in Sri Lanka (P3023)
Authors: Handunnetti Shiroma , Perera Maureen , Herath Nimali , Pathirana Sisira , Mendis Kamini , Premawansa Sunil 
Published: January 2023

Abstract
Plasma levels of pro- and anti-inflammatory cytokines of P. falciparum-infected patients with severe malaria (SM; n=62) and uncomplicated malaria (UM; n=69) from Sri Lanka were assessed. SM patients had significantly higher levels of TNF-α (P&lt;0.01), IL-6 (P&lt;0.01) and IL-10 (P&lt;0.05) compared to the UM patients. Plasma IL-2 levels of these patients were undetectable. TNF-α levels of a third group of patients with uncomplicated P. falciparum malaria, who were recruited during their fever episodes (UMF; n=14) were significantly higher than those of the UM patients (P&lt;0.001) and comparable to SM patients. Plasma IFN-γ levels of SM patients were higher compared to UM patients, but was not statistically significant. Body temperature in both SM and UMF groups were significantly higher compared to UM group whereas percent parasitaemia in all three groups were comparable. Analysis of plasma TNF-α levels and the ratio of TNF-α/IL-10 in UM (n=34) and SM (n=34) patients carrying TNF1 and TNF2 allelic types showed that SM patients carrying TNF2 had significantly higher TNF-α levels as well as TNF-α/IL-10 ratio compared to UM patients carrying TNF1, UM patients carrying TNF2 and SM patients carrying TNF1 (P&lt;0.05). These results suggest that the high circulating TNF-α levels and the inadequate IL-10 response in the SM patients carrying TNF2 allele could have contributed to the development of severe falciparum malarial disease.
Elevated proinflammatory cytokine production by monocytes in HIV(+) individuals at risk for cardiovascular disease (P3033)
Authors: Jalbert Emilie , Parikh Nisha , Seto Todd , Chow Dominic , Shikuma Cecilia , Ndhlovu Lishomwa , Barbour Jason 
Published: January 2023

Abstract
Despite virologic suppression by HIV antiretroviral therapy, residual inflammation associated with chronic HIV infection increases the risk of developing cardiovascular disease. Monocytes have been shown to be major players in the development of atherosclerosis due to their proinflammatory responses to oxidized Low Density Lipoproteins. Our study sought to assess the functional properties of monocytes from peripheral blood of HIV-infected individuals. The cohort consisted of 33 HIV(+) subjects on HAART and 14 HIV(-) risk- and age- matched subjects. Our flow cytometry-based functional assay measured monocyte production of IL-1β, IL-8 and IL-6 in the absence of stimulation and in response to LPS or oxLDL. Without stimulation, HIV(+) subjects had a greater frequency of cells producing IL-1β and IL-8. In the presence of either oxLDL or LPS, both groups increased the frequency of responding cells compared to no stimulation, but HIV(+) subjects maintained a higher frequency of IL-1β(+) and IL-8(+) cells compared to HIV(-). There was no IL-6 production in either group in the absence of stimulation, but upon stimulation with either oxLDL or LPS, there was a higher frequency of IL-6 producing cells in the HIV(+) group. The higher level of inflammatory cytokine production in HIV(+) adults compared to HIV(-), both at rest and in the presence of stimulation, may in part account for increased risk of cardiovascular disease seen in the HIV(+) population.
IL-17 has a critical role to promote protection against Leishmania infantum infection (P3011)
Authors: Nascimento Manuela , Carregaro Vanessa , Lima Djalma , Costa Diego , Dias Fabrício , Ryffel Bernhard , Rossi Marcos , Jesus Amélia , Almeida Roque , Santana João 
Published: January 2023

Abstract
During visceral leishmaniasis the Th1 profile is associated with protection, but the role of IL-17-producing cells remains poorly defined. Our aim was to evaluate whether IL-17 participates in the host immune response against L. infantum and such mechanisms involved. We found that infected humans strongly produce IL-17 during the symptoms peak. Similarly, in murine model IL-17 was induced at the same time of parasitism peak. The IL-17 participates on the parasite control since IL-17R-/- infected mice showed higher parasite burden then WT. In accordance, a lower inflammatory infiltrate was observed in IL-17R-/- mice as a consequence of significant lesser expression of cxcl10, cxcr3, ccl5 and ccr5 in the liver and spleen. Interestingly, an up regulation of ccl17, ccr4 and foxp3 and IL-10 expression was observed in KO mice. Corroborating these data, IL-17 production was potentiated in IL-10-/- infected mice, which were more resistant to infection. Besides, in the absence of IL-17R a smaller expression of IFN-γ, Tbet and iNOS was detected. Indeed, IL-17R-/- infected-macrophages showed a deficiency to kill the parasite and the stimulus with IL-17 and/or IFN-γ in WT infected-macrophage showed that IL-17 acts synergistically with IFN-γ potentiating the NO production and the macrophage leishmanicidal capacity. Together, our results showed that L. infantum triggers IL-17-producing cells that promote the control of parasitism by inducing NO expression and IL-10 modulation.
IL-9-producing cells promote Th1 responses against Mycobacterium tuberculosis. (P3015)
Authors: Alvarez Ivana , Rovetta Ana , Hernández Del Pino Rodrigo , Pasquinelli Virginia , Peña Delfina , Musella Rosa , Palmero Domingo , Chuluyán Héctor , Samten Buka , Barnes Peter , García Verónica 
Published: January 2023

Abstract
IFNγ-secreting cells are critical in the defense against M. tuberculosis (Mtb) infection, but other cells might be required to eradicate the pathogen. IL-9-producing cells were reported to be involved in inflammation and immune diseases, but their role during tuberculosis remains to be clarified. Here we analyzed the involvement of IL-9-secreting cells in the immunity of patients with active disease. We demonstrated that Mtb induced a marked expansion of CD4 IL-9+ cells in peripheral blood from tuberculosis patients. After antigen stimulation, patients with robust Th1 responses to Mtb (high responders) produced higher amounts of IL-9 compared to those with weak Th1 responses (low responders) and a direct significant correlation between IL-9 and IFNγ production against Mtb was detected. To investigate the role of the IL-9 secreted by those lymphocytes, we inhibited IL-9 expression with siRNA, which significantly diminished the production of IFNγ against Mtb. Addition of IL-9 markedly increased the secretion of IFNγ against Mtb, even in low responders. Furthermore, treatment with IL-9 inhibited Mtb-induced apoptosis of IFNγ+ cells, enhancing the survival of the cells. Finally, by studying the effect of neutralizing IL-9 antibodies on polarized Th9 cells, we confirmed that IL-9 participates in the up-regulation of IFNγ production against Mtb. Thus, our results indicate that IL-9 enhances Th1 responses during tuberculosis promoting the cell-mediated immunity to the pathogen.