bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunogenicity of mucosally-administered modified vaccinia Ankara expressing HIV-1 Env protein gp120 in rabbits (VAC8P.1047)
Authors: Jones Dorothy , Pickup David , Staats Herman 
Published: January 2023

Abstract
Modified vaccinia Ankara (MVA) used as a vaccine vector expressing heterologous antigens has been demonstrated to be immunogenic when delivered parenterally while its immunogenicity after mucosal delivery is less defined. To determine the immunogenicity of MVA when administered via different mucosal routes, NZW rabbits were vaccinated on day 0 and 28 with 3 x 108 PFU MVA expressing HIV-1 gp120 by nasal, rectal, or gastric routes. Intramuscular immunization with 1 x 108 PFU of the MVA vaccine was used as the positive control. Serum was collected on days 21 and 42 and antigen-specific antibodies were determined by ELISA. While intramuscular vaccination was the only route that induced a significantly elevated anti-gp120 serum IgG titer, nasal vaccination induced detectable anti-gp120 IgG response on day 42. Immunization by rectal or gastric routes did not induce measurable anti-gp120 IgG responses. Intramuscular and intranasal vaccination, but not gastric or rectal vaccination with MVA, induced significantly elevated serum IgG responses to pox proteins A33R and B5R. We conclude that, under the conditions tested, MVA vaccine vectors are immunogenic when delivered intramuscularly and intranasally but not gastrically or rectally. Insert specific responses after nasal vaccination were not comparable to those achieved by intramuscular vaccination, indicating that use of MVA as a nasal vaccine vector will need to be optimized.
Sendai virus is a safe and immunogenic Jennerian vaccine for human parainfluenza virus-type 1 in 3-6 year old children (VAC8P.1053)
Authors: Penkert Rhiannon , Adderson Elisabeth , Branum Kristen , Sealy Robert , Jones Bart , Surman Sherri , Freiden Pamela , Slobod Karen , Gaur Aditya , Hayden Randall , Allison Kim , Howlett Nanna , Utech Jill , Allay Jim , Knight James , Sleep Susan , Meagher Michael , Russell Charles , Portner Allen , Hurwitz Julia 
Published: January 2023

Abstract
Human parainfluenza viruses are responsible for tens of thousands of hospitalizations and hundreds of thousands of emergency room visits each year in the United States. Included in this group of viruses is human parainfluenza virus-type 1 (hPIV-1), the main causative agent of laryngotracheobronchitis (croup) in young children, for which there is currently no licensed vaccine. In response to this need, we have developed a Jennerian vaccine that employs the use of murine PIV-1 (Sendai virus, SeV) to produce cross-reactive antibodies towards hPIV-1. The vaccine was administered as a single intranasal dose to healthy, hPIV-1 seropositive, 3-6 year old children in a dose escalation study (5 x 105, 5 x 106 or 5 x 107 EID50). Importantly, the vaccine was well tolerated by all participants. No replication of the viral vaccine was detected in the airway of any participant. Within four weeks of vaccination, most participants showed an increase in binding and neutralizing antibodies towards hPIV-1. For several participants, antibody responses remained elevated at six months post-vaccination. The results of this study suggest that hPIV-1 vaccination may be beneficial to boost immunity in seropositive children, and encourage testing of the vaccine in young, seronegative children who are most at risk for respiratory virus disease.
Immunogenicity of inactivated polio vaccine, given with or without mucosal adjuvants dmLT and TCTA1T in rat model (VAC9P.1102)
Authors: park julien , Rho Semi , Kim Jae-Ouk , Song Man Ki , Dey Ayan 
Published: January 2023

Abstract
To eradicate polio, the use of oral polio vaccine (OPV) should be stopped to avoid circulation of vaccine-derived polioviruses (VDPVs). After OPV cessation, transition to inactivated polio vaccine (IPV) is recommended. This can only be accomplished when IPV induce better mucosal protection. This can be achieved by adding mucosal adjuvant to the IPV formulation. In the present study we used wistar rats to elucidate the adjuvant effect of double mutant heat-labile toxin (dmLT) of E. coli and recombinant cholera toxin (TCTA1T) on IPV. Rats were administered by intramuscular injection of 1.0-0.6-1.0 D-antigen units (DU) of Inactivated Polio type 1, 2 and 3 /rat. The E. coli dmLT and TCTA1T was used as adjuvant and was administered subcutaneously along with IPV. The control groups received IPV alone or PBS. Rats were immunized on days 0, 14 and 28 days. After last vaccination mice were sacrificed and spleen cells and blood were collected. IgA secreting cells specific for Polio Type1, 2 and 3 were measured by ELISPOT. Intramuscular immunization results in robust mucosal response in inactivated polio antigens + dmLT group compared to other groups. Further measurement of antibodies in serum doesn’t show significant difference among IPV and adjuvanted IPV groups. Thus the present data suggest that vaccination with IPV and dmLT results in specific induction of mucosal immunity, which will provide barrier to infection from oral fecal route.
Identification of MHC bound peptides from murine bone marrow derived dendritic cells infected with Salmonella (VAC9P.1108)
Authors: Karunakaran Karuna , Yu Hong , Jiang Xiaozhou , Chan Queenie , Goldberg Michael , Foster Leonard , Jenkins Marc , Brunham Robert 
Published: January 2023

Abstract
Salmonella infections cause a spectrum of diseases ranging from self-limiting diarrhea to life-threatening systemic disease. Diseases caused by Salmonella are treatable with antibiotics but successful antibiotic treatment has become increasingly difficult due to antimicrobial resistance. An effective vaccine is therefore the best strategy to reduce these diseases. A clear understanding of host immunity to Salmonella is essential to aid in the development of a vaccine. Protective immunity against Salmonella depends on a wide range of innate and adaptive immune mechanisms and T cell-mediated immune responses are important in the host control of intracellular Salmonella infection. Our laboratory used an immunoproteomics approach to identify Chlamydia T cell antigens that exhibited significant protection against Chlamydia infection in mice when used as vaccines. These results demonstrate that T cell antigens identified by immunoproteomics can be successfully exploited as T cell vaccines against an intracellular pathogen. Recently we generated dendritic cells from bone marrow of C57BL/6 mice and infected with Salmonella enterica SL1344 followed by elution of MHC class I and class II-bound peptides. The sequences of the purified peptides were then identified using tandem mass spectrometry. We identified 87 MHC class II and 23 MHC class I Salmonella derived peptides. These antigens are of use in Salmonella immunobiology research and as potential Salmonella vaccine candidates.
Effect of anticoagulants on the measurement of cholera vaccine-induced humoral immune responses (VAC9P.1101)
Authors: Yang Jae Seung , kang Seok Seong , Yun Cheol-Heui , Han Seung Hyun 
Published: January 2023

Abstract
Anticoagulated blood is widely used for the simultaneous evaluation of humoral and cellular immune responses after the vaccination. However, anticoagulants in the plasma may interfere with the quantitative and qualitative analyses of antibodies. In the present study, we examined the potential interference of the widely-used anticoagulants including heparin, acid citrate dextrose (ACD), and ethylenediaminetetraacetic acid (EDTA) on bactericidal activity against V. cholerae and its LPS-specific IgG, IgM, and IgA levels in sera and plasma samples from cholera patients or vaccinees. Serum vibriocidal activity was inhibited in the presence of EDTA and ACD whereas no change was found when heparin was used. Moreover, vibriocidal titer of plasma, obtained from vaccinees using tube containing heparin, was 100% identical with the titer of corresponding sera while 37.5%, 50% in the plasma containing EDTA and ACD were matched, respectively. Pearson correlation coefficient (r) for LPS-specific IgG, IgM, and IgA between serum and heparinized plasma were 0.997, 0.945, and 0.716, respectively. Collectively, heparin is suggested to be an appropriate anticoagulant for collection of blood for measuring vibriocidal activity and antibody in the plasma after cholera vaccination.
Genetic markers for resilience and susceptibility to Johne's disease In red deer (Cervus elaphus) (VET1P.1120)
Authors: Brennan Liam , O'Brien Rory , Griffin Frank 
Published: January 2023

Abstract
Mycobacterium avium subspecies paraturberculosis is the causative agent of Johne’s disease, an enteritis infection that affects ruminants.This disease has been studied in deer, but there is little information regarding immunity or disease immunopathology. There is evidence that host genetics contribute to resilience or susceptibility to the disease. This study sought to identify genes in the innate and adaptive immune systems as biomarkers for resilient (R) or susceptible (S) phenotypes in red deer. We have bred over 60 animals from sires with a confirmed R or S phenotype. Animals were bled routinely and monocyte derived macrophages (MDMs) or whole PBMCs were obtained and cultured in vitro. Gene expression was determined using qPCR. The 15 most informative gene targets were chosen from a panel of 60 using GenEx software MDMs showed higher levels of expression of immune genes such as IL1A, TNFA, IL23A and IL12A in S animals, as well as genes not typically associated with the immune system, such as PKLR that exhibit this pattern. Conversely, PBMC samples from these animals exhibit higher levels of expression of immune genes such as IFNY, IL17A and IL2 in R animals. These results have been reproduced in animals bred from different sires over several years. We intend to incorporate these gene markers into diagnostic tests to identify R and S animals at an early age. These genes may also provide deeper insights into immune parameters that determine disease outcome in an animal.
Resistin-like molecules reduce influenza A virus morbidity in mice (VIR5P.1158)
Authors: Samarasinghe Amali , Lin Yanyan , LeMessurier Kim , Woolard Stacie , McCullers Jonathan 
Published: January 2023

Abstract
Resistin-like molecules (RELMs) are recently discovered cysteine rich secreted proteins abundant in a variety of organs including the lungs. RELMα has an inhibitory role in the lungs by downregulating inflammation, while RELMβ promotes inflammation. We noted a markedly greater expression of RELMs in the lungs of mice with acute asthma and influenza, which were protected against severe influenza morbidity and hypothesized that RELMα and RELMβ proteins enhance anti-viral defenses. Recombinant RELMα and RELMβ proteins were administered intranasally to C57BL/6 mice starting three days prior to viral infection and continued on a daily basis following viral infection. Mice were infected with 1000 TCID50/ml of A/CA/04/2009 pandemic influenza A virus. Animals were sacrificed on days 3, 5, 7, and 9 following viral infection for sample acquisition. Animals not treated with exogenous RELMs served as controls. Administration of RELM proteins led to a reduction in airway resistance after influenza virus infection. Inflammation in the airways increased in RELM-treated mice with macrophages and neutrophils taking precedence and a prominent increase in CD8+ T cells at late timepoints. A significant reduction in viral burden occurred in the lungs of RELM treated mice. The administration of RELM proteins directly impacted the host immune response against influenza A virus. Future studies will investigate the mechanisms by which RELMs mediate protection against influenza.
Characterizing the T cell receptor clonotype repertoire of an atypical HLA class II-restricted CD8 T cell response in HIV-1 infection (VIR6P.1170)
Authors: Lamothe Pedro , Ranasinghe Srinika , Crawford Frances , White Janice , Clayton Gina , Power Karen , Garcia-Beltran Wilfredo , Allen Todd , Shalek Alex , Kappler John , Walker Bruce 
Published: January 2023

Abstract
CD8 T cells targeting peptides presented by human leukocyte antigen (HLA) class II are atypical. Little is known about how the CD8 T cell receptor (TCR) recognizes peptides on HLA class II. We analyzed the TCR repertoire of a class II-restricted CD8 T cells targeting a HIV peptide. We identified an atypical HIV-specific CD8 response to DV16 peptide on HLA-DRB1*11. We sequenced the TCR of the class II-tetramer sorted cells. To measure the binding kinetics of the TCR with the peptide-MHC we used surface plasmon resonance (SPR). Analysis of 68 sequences showed that TCR-beta repertoire has a single TRBV2*01 clonotype. We looked at 64 sequences of TCRalpha and found that the repertoire had two clonotypes: TRAV26-1*02 and TRAV6*02. SPR revealed that only the TRAV6 was able to bind to the peptide-MHC. We studied 27 sequences of the CD4 T cells targeting the same MHC class II-peptide. We observed a polyclonal response of 16 clonotypes. 13 clonotypes used the same TRBV2*01 gene but had different rearrangements. Interestingly, two of these sequences are exactly the same as the dominant clonotype from the CD8 response. We found that the TCR is shared between CD8 and CD4 T cells targeting the same class II HLA-peptide. These data suggest that atypical CD8 cells express two different TCR alpha, possibly due to inefficient allelic exclusion during development. The use of the same TRBV2*01 by different CD4 clonotypes may suggest an atypical docking of TCR in the binding with peptide-HLA.
Characterization of functional antiviral CD8 T cell immune responses of HIV elite controllers by high dimensional phenotyping using mass cytometry (VIR6P.1169)
Authors: Kleinsteuber Katja , Rashidi Narges , Soghoian Damien , Nchinda Nzuekoh , Mavrothalassitis Orestes , Buus Soren , George Tad , Ornatsky Olga , Walker Bruce 
Published: January 2023

Abstract
Elite controllers (EC) are individuals who control HIV infection without therapy through CD8 T cells. Chronic infections can lead to T cell exhaustion characterized by reduced proliferation and cytokine expression as well as upregulation of co-inhibitory molecules like PD-1. CD8 T cells from EC are more effective at killing HIV-infected targets than those from progressors. The underlying mechanism is not completely understood, but is affected by HLA type, T-cell receptor clonotype, viral epitopes targeted, and likely also by a complex regulatory network of activation and inhibition. However, the particular phenotype of activation and exhaustion markers associated with the increased killing of HIV-specific CD8 T cells from EC has remained unclear. To elucidate these regulatory networks, we performed high-dimensional phenotyping of HIV-specific CD8 T cells using mass cytometry (CyTOF), a cutting edge technology combining flow cytometry and mass spectrometry that enables analysis of up to 40 markers with minimal signal overlap. We established and validated a CyTOF panel of 33 markers to identify up to 6 HIV-specific CD8 T-cell subsets using tetramer technology and characterize memory, activation and exhaustion status of those cells. Use of our panel enables the in-depth characterization of HIV-specific CD8 T cells at a level of unprecedented granularity and may reveal signatures of activation and exhaustion networks that underlie the enhanced lytic capacity of these cells in EC.
A framework for unbiased, robust and system-wide characterization of MHC-bound peptides and epitopes (APP5P.111)
Authors: Foster Leonard , Chan Queenie , Kuan Charlie , Yu Hong Bing 
Published: January 2023

Abstract
Vaccines are the most inexpensive medicine in the long run, yet developing them remains a slow process. A major challenge is to identify proteins efficiently processed by the host and presented on major histocompatibility complexes (MHCs). Furthermore, the extreme polymorphism of the MHCs means that candidate antigens must be tested across many genotypes. Immunopeptidomics, the study of all peptides presented by the host’s immune system, holds promise but is limited by informatics:standard approaches result in high error rates for these samples. We describe here an experimentally validated informatic treatment of mass spectrometric data from peptides eluted from the surface of antigen-presenting cells that can identify epitopes bound by specific MHCs, predict their core binding regions, and reveal consensus binding motifs of MHC alleles. Our approach avoids the biases inherent with MHC immunopurification or prior consensus motifs and enables rational vaccine design by allowing rapid and sensitive screening of individual immunopeptidomes.
Determinants of immunogenicity: flogging the old ovalbumin, again (APP5P.116)
Authors: Karandikar Sukrut , Srivastava Pramod 
Published: January 2023

Abstract
During antigen presentation, the products of intracellular proteolysis capable of binding MHC I are presented to T cells on the cell surface in context of MHC I. We queried the sequence of ovalbumin (OVA) with the NetMHC 3.0 algorithm and based on a suitable NetMHC Matrix score or predicted IC50 values, identified 19 putative epitopes with potential to bind H-2Kb or H-2Db. Generally, IC50 of 500 nM is used as a threshold to distinguish between candidate and non-candidate epitopes; sequences with an IC50 &lt;=500 nM are considered to be good binders and candidate epitopes (Assarsson et al. J Immunol. 2007). We observe that 10 of the 19 putative epitopes of OVA have measured IC50 &lt;= 500 (range: 1.5 nM - 248 nM). The remaining nine putative epitopes are weak binders (IC50 &gt; 500; range: 928 nM- 13219 nM). Interestingly, three of the nine weak binders elicit antigen specific responses in C57BL/6 mice. Six of the ten strong binders also elicit such responses, while four do not. Altogether, we have identified nine novel MHC class I restricted epitopes of ovalbumin, one of the most exhaustively used model antigens. Finally, we observe that synthetic peptides corresponding to two epitopes of OVA (one weak binder and one strong binder) do not elicit specific responses upon immunization; at the same time, responses against these epitopes can be observed upon immunization with OVA. We are exploring the molecular mechanisms of these observations.
The effect of HLA-DM on cooperativity of peptide binding to MHC class II (APP5P.107)
Authors: Templeton Megan , Ferrante Andrea 
Published: January 2023

Abstract
HLA-DM (DM) plays a critical role in generating the peptide-MHC class II (MHCII) repertoire expressed on the surface of Antigen Presenting Cells and recognized by CD4+ T Cells. The action of DM is pH-dependent, and its effect increases at acid conditions. Previous studies performed at pH 7.4 in the absence of DM have shown that peptide binding to and release from MHCII is a flexible process as evidenced by cooperativity. Cooperativity indicates that peptide binding does not rely on independent contributions of single-point interactions, instead multiple interactions along the peptide-binding groove determine binding in a non-additive fashion To resolve whether cooperativity of binding, thus system flexibility, is a function of DM activity, we measured the affinity of peptides derived from influenza H3 HA306-319 via cycle-mutation for the human MHCII allele HLA-DR1 (DR1) at pH 5.4 and 6.4. Through competitive binding assays and peptide release studies we have determined that DM reduces cooperativity in peptide binding and peptide-MHCII kinetic stability, and the effect is a function of pH conditions. These results suggest that one component of DM action during peptide exchange is a restraining of complex structural flexibility, which imposes a higher energetic threshold for stable binding.
A critical role of Th17 cells in the development of murine Sjogeren’s syndrome (BA11P.132)
Authors: Lin Xiang 
Published: January 2023

Abstract
Sjögren’s syndrome (SS) is characterized by progressive inflammation and tissue damage of salivary glands (SG) and lacrimal glands, leading to dry-mouth and dry-eye symptoms as a result of reduced saliva and tears secretion. Histopathologically, CD4+ T cells were the major population in the infiltration and mediated tissue destruction. However, a role of IL-17-producing CD4+ T cell (Th17 cell) has not yet been elucidated in SS pathogenesis. In the present study, experimental SS was induced in normal mice by immunization with proteins extracted from SG. A rapid increase of Th17 cells was observed in the spleen and draining cervical lymph node at acute stage. Interestingly, this pattern also emerged in the draining lymph node of non-obese diabetic mice, another murine model of SS. Further, SG-infiltrating Th17 cells were detected and increased along disease progression. In IL-17 deficient (IL-17-/-) mice, no apparent salivary dysfunction and autoantibodies were observed upon immunization. However, upon adoptive transfer of Th17 cells from wild type mice, high titer of autoantibodies, salivary dysfunction with lymphocytic infiltration were fully restored in IL-17-/- mice. Together, our findings have demonstrated a critical role of Th17 cells in SS pathogenesis.
Specifying the autoantibody-inducing CD4 T cell causing systemic lupus erythematosus out of PD-1+CD45RBlo122lo CD4 subpopulation using mass spectrometry (BA11P.135)
Authors: Shiozawa Shunichi , Miyazaki Yumi , Tsumiyama Ken 
Published: January 2023

Abstract
Objective: We have shown that repeated immunization of mice with antigen reproducibly led to development of experimental systemic lupus erythematosus (SLE), where a novel T cell type which we term an autoantibody-inducing CD4 T (aiCD4 T) cell was generated via TCR revision at periphery. The aiCD4 T cell induces varieties of autoantibodies and also helps full maturation of CTL to induce lupus tissue injuries. We have previously identified that aiCD4 T cell belongs to PD1+CD45RBlo122lo CD4 T subpopulation. Here we tried to further specify the aiCD4 T cell by membrane surface markers. Methods: BALB/c mice were repeatedly 12x immunized with OVA. To isolate aiCD4 T cell, we extracted cytoplasmic and membrane fractions of PD1+CD45RBlo122lo CD4 T subpopulation. They were electrophoresed and protein fractions unique to the PD1+CD45RBlo122lo CD4 T subpopulation were identified thru mass spectrometry. Results: Candidate membrane markers as suggested by mass spectrometry were DNA topoisomerase 2-beta (181 kDa), DOCK 8 (238 kDa), VWA5A (87 kDa), STAT5b (89 kDa), epidermal growth factor receptor substrate 15-like 1 (99 kDa) and FLAP (18 kDa) for aiCD4 T cell, among which we found that DOCK 8 was significantly increased in the membrane fraction of PD1+CD45RBlo122lo CD4 T subpopulation. Conclusion: We identified DOCK 8 as a possible candidate membrane marker characterizing the aiCD4 T cell as found within PD-1+CD45RBlo122lo CD4 T subpopulation.