bluuebunny/crossref_metadata_2025_split huggingface dataset:
NK Cell Mediated Contact Sensitivity is Elicitable 1 Hour After Immunization and Depends on IFN-gamma and IL-12 Production (B101)
Authors: Askenase Philip W , Szczepanik Marian , Majewska Monika 
Published: January 2023

Abstract
NK cell mediated contact sensitivity (CS) has recently been described. We confirmed CS in SCID and RAG1−/− mice but not in SCID-beige mice without NK cell function. NK mediated CS was transferred with liver mononuclear cells and unlike conventional CS not with mixed lymph node and spleen cells. Positive selection with anti-DX5 beads confirmed the NK nature of the transferring cells. Participation of NKT cells and B-1 cells was ruled out with Jα18−/− and JH−/− mice. Remarkably, NK CS was elicitable 1-hr after immunization and with an early 2-hr component 1 day after LMNC transfer, and lasted at least 7 days. Adoptive cell transfer required MHC matching and IL-12 and IFN-γ−/− were involved in the mechanism of CS.
In conclusion, NK CS is a distinct entity. IL-12 and IFN-γ are involved in the effector mechanism of NK-mediated CS, similar to classical CS mediated by Th1 cells. The requirement for MHC matching may involve the specificity of the NK receptor, which in this case may be hapten altered self. In conclusion, NK cells provide a very rapid and long lasting, Ag-specific mechanism to elicit cell-mediated immunity, with features of innate and acquired immunity.
Supported by NIH grants AI-59801 and AI-07174 to PWA; and the Polish Committee of Scientific Research to MS.
Streptococcus pyogenes activates human plasmacytoid and myeloid dendritic cells (B123)
Authors: Veckman Ville , Julkunen Ilkka 
Published: January 2023

Abstract
Human peripheral blood contains two major dendritic cell populations, namely CD11c−CD123+ plasmacytoid DCs (PDCs) and CD11c+CD123− myeloid DCs (MDCs). Here we have compared PDC and MDC activation in response to Gram-positive Streptococcus pyogenes (group A streptococci) and influenza A virus. S. pyogenes stimulation induced the maturation of both DC types as evidenced by enhanced expression of co-stimulatory molecules and production of pro-inflammatory cytokines and chemokines. Furthermore, S. pyogenes-stimulated PDCs and MDCs activated naive CD4+ T cells and enhanced their Th1 type cytokine production. Influenza A virus infection induced rapid PDC activation whereas MDCs were extremely sensitive to influenza A virus-induced cell death. The most significant differences between DC types were seen in the production of IL-10 and IL-12, which were produced only by S. pyogenes-stimulated MDCs. Although S. pyogenes was able to induce PDC activation, only influenza A virus infection resulted in detectable IFN-α production. Our results show that depending on the infecting microbe the functions of PDCs and MDCs may be partially overlapping suggesting a considerable flexibility of the human DC system.
Generation of a protective T cell response following viral infection of the CNS is independent of either IL-12 or IL-23 (B141)
Authors: Held Katherine , Glass William , Orlovsky Yevgeniya , Shamberger Kimberly , Petley Ted , Branigan Patrick , Cunningham Mark , Benson Jacqueline , Lane Thomas 
Published: January 2023

Abstract
The functional role of IL-12 and IL-23 in generating effector T cells following viral infection of the central nervous system (CNS) was determined. Instillation of mouse hepatitis virus (MHV) into the CNS of C57BL/6 mice results in an acute encephalitis followed by a chronic demyelinating disease. MHV-infected mice express mRNA transcripts specific for IL-12p35 and IL-23p19 subunits within the brains. Antibody-mediated blocking of IL-23 (anti-IL-23p19) or IL-12 and IL-23 (anti-IL-12/23p40) did not reduce numbers of virus-specific T cells and treated mice cleared virus from the CNS demonstrating that blocking IL-23 does not affect the generation of a Th1 response. Therapeutic administration of anti-IL-23p19 to infected mice with established demyelination did not attenuate disease indicating IL-23 does not contribute to viral-induced demyelination. Treatment with anti-IL-12/23p40 or anti-IL-23p19 resulted in inhibition of EAE, an autoimmune model of demyelination highlighting the importance of IL-23 in CNS autoimmune inflammation and disease. These data indicate that i) IL-12 and IL-23 signaling are dispensable in generating a protective response following CNS infection with MHV and ii) IL-23 amplifies disease in a model of autoimmune demyelination. Therefore, targeting of IL-23 for the treatment of autoimmune diseases may offer unique advantages by reducing disease severity without muting protective responses following viral infection.
Modulation of Adaptive Immune Responses in Chickens by Low Pathogenicity Avian Influenza Virus H9N2 (B142)
Authors: Xing Zheng , Cardona Carol J , Li Jinling , Tran Tu , Andrada Jason 
Published: January 2023

Abstract
Most low pathogenicity avian influenza (LPAI) viruses cause no or mild diseases in avian species. The H9N2 subtype are commonly detected LPAI viruses that have crossed the species barrier to infect humans, unlike most other LPAI viruses. We report in this study that chicken macrophages are susceptible to infection with H9N2 and H6N2 viruses and infection led to apoptosis. Additionally, in H9N2 virus-infected macrophages, major histocompatibility complex (MHC) antigens, class II in particular, were downregulated, which also occurred in the lungs of H9N2 infected chickens. Pro-inflammatory cytokines and chemokines were differentially regulated in both infected macrophages and lungs. Among the regulated genes, IL-4, IL-4R, and CD74 were downregulated, all of which are pivotal in the activation of CD4+ helper T cells and humoral immunity. Remarkably, in H9N2 virus-infected chickens the antibody response was largely suppressed. This is in contrast to what was observed in those infected with H6N2 virus. This finding suggests that the pathogenesis of H9N2 infection in chickens may involve humoral immune suppression. Modulation of MHC antigens and the subsequent impact on the immune responses in mammals infected with avian influenza viruses is largely unclear. These findings raise questions about how some LPAI viruses regulate avian immune responses and if they have similar effects on mammalian immune function.
Pgp (MDR1)-dependent resisrance of malignant cells towards natural killer (NK) cells is reversed by IL-2 and by the Pgp inhibitor KT-5720: implications on graft versus tumor (GVT) effects in adoptive immunotherapy (B144)
Authors: Nagler Arnon , Bar Iris , Ostrovsky Olga , Galski Hanan 
Published: January 2023

Abstract
P-glycoprotein (Pgp) encoded by the MDR1 gene, may render tumor resistant to NK and CTLs mediated immunotherapy (DLI). We assessed Pgp expression on tumors and their response to DLI & tried to reverse tumor resistance towards NK cells.
The KB-V1, Pgp over-expressing cell line was found significantly less sensitive to resting & IL-2 activated NK cells than its parental KB-3-1, Pgp non expressing cell line. NK-resistant Colo205 cell line expressed high level of Pgp RNA, while low expression level was found in NK-sensitive K562 cell line. The allele polymorphism C3435T of MDR1 is associated with phenotypic variations in Pgp expression, with high-, intermediate- or low-expression level of the CC, CT and TT genotypes, respectively. Both KB-V1 & Colo205 (MDR1 over-expressing and NK-resistant) cell lines carried the CC genotype while the K562 cell line (MDR1 low-expressing, and NK-sensitive) carried the TT genotype. Incubation of resting NK cells with IL-2 increases significantly the killing of both KB-3-1 & KB-V1 cell lines. The Pgp inhibitor KT-5720 increases the killing of KB-V1 cells (but not of KB-3-1 cells) by resting NK cells. In summary:
Low cytolytic response of tumor cells to NK-cells is associated with high Pgp expression and the CC genotype of the MDR1 geneIL-2 & the Pgp inhibitor, KT-5720, reverse Pgp-dependent resistance of tumors towards NK cellsExposure of NK-cells to IL-2 increases their cytolytic activity. The effect of IL-2 is larger on Pgp-high expressing- than on Pgp-non expressing-target cellsSince KT-5720 is a novel Pgp inhibitor this study may lead to future clinical application of KT-5720, to increase GVT effects in adoptive immunotherapy.
Memory T cells in the tumor-infiltrating lymphocytes for adoptive cell transfer therapy (B145)
Authors: Zhou Juhua , Nagarkatti Prakash , Robbins Paul F , Rosenberg Steven A , Nagarkatti Mitzi 
Published: January 2023

Abstract
It has been documented that immune memory has a critical role in mediating protection against infections as well as potentiating allergic and autoimmune diseases. Memory T cell populations in human anti-tumor tumor-infiltrating lymphocytes (TILs) for adoptive cell transfer have not fully been characterized. FACS analysis showed that T cells in the initial TIL microcultures generated from the tumor tissues of melanoma patients did not express CCR7, however, 67¡À5 % of T cells expressed CD27, 70¡À6 % of T cells expressed CD28 and 35¡À5 % of T cells expressed CD62L. As compared with the TIL microcultures before large-scale expansion, T cells in the administered TIL samples had significantly decreased levels of CD62L and CD27 expression. CD28 expression, however, did not change dramatically after large-scale expansion. These memory phenotypic T cells also expressed high levels of CD44. Furthermore, T cell expansion significantly decreased the percentage of CD8+CD44+ T cells. These results indicated that effector memory T cells were present in TIL samples and the maintenance of memory T cells in TILs may represent an important factor in effective adoptive cell transfer therapy.
(Supported by NIH grants R01AI053703, R01ES09098, R01 AI058300, R01DA016545, R01HL058641 and the Intramural Research Program of the NIH, National Cancer Institute, Center for Cancer Research.)±±
OX40 signals preferentially mediate the generation of effector memory but not central memory CD4+ T cells (B15)
Authors: Soroosh Pejman , Ine Shouji , Sugamura Kazuo , Ishii Naoto 
Published: January 2023

Abstract
Memory T cells can be divided into effector memory (TEM) and central memory (TCM) subsets based on their effector function and homing characteristics. Although previous studies have demonstrated that TCR and cytokine signals mediate the generation of memory T cells, it is unclear how the generation of the two memory subsets is regulated. We found that OX40-deficient mice showed a marked reduction in the number of CD4+ TEM cells, while the number of CD4+ TCM cells was normal. Adoptive transfer experiments using Ag-specific CD4+ T cells revealed that OX40 signals during the priming phase were indispensable for the optimal generation of the CD4+ TEM, but not the CD4+ TCM population. Furthermore, CD4+TEM cells that were generated in the absence of OX40 signals showed impaired cytokine production. Collectively, the present results indicate that the generation of functional CD4+TEM cells, but not TCM cells, is critically regulated by OX40 signals.
Defective CCR7 function in aging: implications for dendritic cell migration (B156)
Authors: Grolleau-Julius Annabelle , Harning Erin , Yung Raymond 
Published: January 2023

Abstract
We recently reported that bone marrow-derived dendritic cells (DCs) from old C57BL/6 mice are less effective than their young counterpart in inducing the regression of pre-established B16-OVA melanomas. However, the mechanisms for this are unclear. DCs migration to lymphoid organs is essential to effective DC anti-tumor activity and is regulated by interactions between CCR7 and its ligands CCL21 and CCL19. We therefore examined the effect of aging on DC migration, CCR7 expression and function. Young (3–6 months) and old (18–20 months) DCs were labeled with CFSE and CMPTX, and their in vivo migration quantified by fluorescent microscopy. Half as many aged DCs were present in the popliteal lymph nodes (p&lt;0.025). DCs from aged mice express 50% less CCR7 compared to young DCs (p&lt; 0.0005). Surprisingly, the reduced gene expression did not result in any significant change in CCR7 surface expression on DCs. Nevertheless, the reduced CCR7 gene expression correlated with the impaired in vitro migration of aged DCs in response to CCL21 (p&lt; 0.05). Finally, we showed that aged DCs have impaired signaling following exposure to CCL21. In conclusion, our data showed that old DCs have defective in vivo migration that correlates with reduced CCR7 gene expression, as well as impaired in vitro CCL21 migratory response and signaling. These findings may contribute to the observed impaired DC tumor immunotherapeutic response in aging.
Ex vivo expansion of PR1-CTL elicited by autologous dendritic cells pulsed with PR1 peptide for adoptive immunotherapy of leukemia (B157)
Authors: Wang Changqing , Ma Qing , Wieder Eric , Lu Sijie , Molldrem Jeffrey J. 
Published: January 2023

Abstract
Cytotoxic T lymphocytes (CTL) specific for PR1, a HLA-A*0201-restricted peptide derived from proteinase 3, are capable of killing leukemia cells and contribute to the elimination of CML. Cytogenetic remission of CML after treatment with IFNα correlates with an expansion of PR1-CTL in patients. INFα is also an abundant cytokine produced by plasmacytoid DC which is critical for priming adaptive immune responses and MHC class I antigen cross-presentation. To determine whether a clinical trial with adoptively transferred PR1-CTL will benefit leukemia patients, we used INFα in combination with GM-CSF for ex vivo expansion of PR1-CTL and examined whether theses PR1-CTL can eliminate AML in the established NOD/SCID model. PBMC from HLA-A*0201+health donors were stimulated weekly with autologous DC pulsed with PR1 peptide in the presence of INFα and GM-CSF. By 3 weeks, there were 38.7% of PR1-CTL in the culture compared to 1.83% detected in the original sample using PR1-tetramer staining. We found that PR1-CTL generated in vitro can migrate to sites of disease and maintain their capacity to eliminate the human primary AML cells in NOD/SCID mice. Mice received AML alone had 72–88% blasts in a hypercellular marrow, whereas mice that received AML plus PR1-CTL had normal hematopoietic elements and only 10–11% blasts in a hypocellular marrow. We conclude that ex vivo expansion of PR1-CTL using this method is suitable for adoptive immunotherapy of AML.
Differential Type I Interferon Induction by Respiratory Viruses (B179)
Authors: Durbin Joan E. , Jewell Nancy A. , Vaghefi Negin , Mertz Sara E. , Akter Parvis , Peebles R. Stokes , Bakaletz Lauren O , Durbin Russell K. , Flano Emilio 
Published: January 2023

Abstract
Type I interferon (IFN) induction is an immediate response to virus infection, and very high levels of these cytokines are produced when the TLRs expressed at high levels by plasmacytoid dendritic cells (pDCs) are triggered by viral nucleic acids. Unlike many RNA viruses, respiratory syncytial virus (RSV) does not appear to activate pDCs through their TLRs, and it is not clear how this property impacts IFN-alpha/beta induction by this virus in vivo. In this study we compared type I IFN production by RSV and influenza A virus infection of BALB/c mice and found that while both viruses induced IFN-alpha/beta production by pDCs in vitro, only influenza virus infection could stimulate type I IFN synthesis by pDCs in vivo. In situ hybridization studies demonstrated that the infected respiratory epithelium was a major source of IFN-alpha/beta in response to either virus; but in pDC-depleted animals only type I IFN induction by influenza virus infection was impaired.
This work was supported by grants R01AI47226 and R01DC006468 to JED from the National Institutes of Health.
MKP-1, a critical regulator of the innate immune response to Gram-negative bacterial infection (B180)
Authors: Frazier Joshua , Wang Xianxi , Kuhlman Joshua , Liu Yusen 
Published: January 2023

Abstract
Mitogen-activated protein (MAP) kinases play a critical role in the innate immune response by inducing the transcription of pro-inflammatory cytokines. MAP kinase phosphatase (MKP)-1 is an archetypal member of the MKP family which inactivates p38 and JNK MAP kinases. We have previously shown that deficiency of MKP-1 predisposes experimental animals to mortality upon challenge with lipopolysaccharide, a Gram-negative bacterial endotoxin. However, the impact of MKP-1 deficiency in the setting of true Gram-negative bacterial infection has not been elucidated. We hypothesize that MKP-1 deficient animals will experience increased mortality from live bacterial infection compared to wild type animals. Age- and sex-matched wild type and MKP-1 deficient mice were injected intravenously with Escherichia coli at a dose of 2.5 x 107cfu/g body weight. After treatment, the animals were monitored for 48 h and mortality data collected for analysis. The mortality of the MKP-1 deficient animals was significantly higher than that of similarly treated wild type mice (p = 0.032). Our results indicate that MKP-1 deficiency confers a disadvantage to animals facing live Gram-negative infection. Thus, MKP-1 appears to play a critical role in down-regulating the inflammatory response to Gram-negative bacterial pathogens.
Activation of C5 via the lectin pathway of complement (B181)
Authors: Salvi Veena P. , Rajagopalan Rema , Kunkalla Kranthi , Rawal Nenoo 
Published: January 2023

Abstract
Activation of C5 via the lectin pathway of complement was analyzed by examining C5 cleaving properties of lectin pathway C3/C5 convertases. C4b-dependent C3/C5 convertase, ZymM1,C4b,C2a and EManM1C4b,C2a (M1 represents MBL-MASPs) assembled on zymosan or sheep erythrocytes coated with mannan (EMan) exhibited a weak affinity for C5 (Km = 2.7–5.3 μM). Very high affinity C5 convertases (ZymM1,C3bC4b,C2a and EManM1,C3bC4b,C2a) were generated when low affinity C4b-dependent C3/C5 convertases were allowed to deposit C3b by cleaving native C3. ZymM1,C3bC4b,C2a and EManM1,C3bC4b,C2a exhibited Km (4.9–8.3 nM) well below the normal concentration of C5 in blood (0.37 μM). Although, the Km, Vmax, and kcat of C3/C5 convertases of lectin pathway were similar to those reported for classical pathway, the ability of C4b to bind C2 indicated that every C4b deposited on zymosan/EMan was capable of forming a convertase. The findings differ from those reported for classical pathway C3/C5 convertase where one out of four C4b molecules deposited formed an enzyme. The four times greater potential of the lectin pathway than the classical pathway in generating C3/C5 convertases and producing proinflammatory products, such as C3a, C4a, and C5a implies that activation of complement via the lectin pathway might be a more prominent contributor to the pathology of inflammatory reactions.
Research supported by NIH grant HL073804
Differential regulation of cytotoxicity and cytokine production via surface receptors in CD45-deficient natural killer cells (B185)
Authors: Hesslein David G.T. , Takaki Rayna , Hermiston Michelle L. , Weiss Arthur , Lanier Lewis L. 
Published: January 2023

Abstract
Little is known about how CD45 regulates NK cell receptor signaling and the resulting effector functions. We find that NK cells from CD45-deficient mice are competent for immunoreceptor tyrosine-based activation motif (ITAM)-dependent receptor-induced cell-mediated cytotoxicity. In contrast, CD45−/− NK cells are completely deficient for cytokine secretion after stimulation with ligands to or antibodies against the ITAM-associated receptors, NK1.1, CD16, Ly49H, Ly49D, and NKG2D. This deficiency in cytokine/chemokine production occurs at the level of mRNA expression. After receptor engagement, the extracellular regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) activation was markedly perturbed, whereas p38 activation was not substantially affected. Further more, we have uncovered other cytotoxicity pathways that are defective in CD45−/− NK cells. These findings indicate that CD45 is not required for ITAM-dependent cytolytic activity, but is required for cytolytic activity through other receptor pathways and for ITAM-dependent cytokine production.
Dry powder formulations of plague F1-V vaccine provide protective immunity in mice (B195)
Authors: Huang Joanne , Ferriter Matthew S. , Alarcon Jason B. , Mikszta John A. , Evans Michelle , D’Souza Ajit , Ford Brandi , Stewart Todd , Amemiya Kei , Ulrich Robert G , Sullivan Vince J. 
Published: January 2023

Abstract
The potential use of Y. pestis as a bioterror agent is a great concern. Development of a more stable powder vaccine against Y. pestis and using non- or minimally- invasive administration methods could provide an effective alternative to traditional intramuscular (IM) injection of liquid formulations.
We evaluated two powder formulations of a recombinant F1-V fusion protein of Y. pestis for vaccination against plaque in a mouse model. Mice were immunized with reconstituted F1-V powders via IM, microneedle-based intradermal (ID) or non-invasive intranasal (IN) administration. By standard IM injection, the reconstituted powders induced serum antibody responses and provided protection against 1000 LD50 Y. pestis lethal subcutaneous challenge at levels equivalent to those elicited by unprocessed liquid vaccine (70 – 90% protection). Overall, ID delivery showed similar efficacy as IM injection, while IN delivery required an extra dose of vaccine to achieve a level of protection similar to that provided by injection.
In summary, we demonstrate the feasibility of using reconstituted F1-V powders and microneedle-based ID delivery to provide protection against plague in a mouse model. Nasal delivery, while feasible, was less effective than injection in this study. The potential use of these alternative delivery methods and powder vaccine formulations may result in substantial health and economic benefits.
Abrogation of potent humoral vaccine responses in patients on long-term anti-CD25 therapy (B196)
Authors: Ragheb Jack A , Buggage Ronald , Reed George , Levy-Clarke Grace , Nussenblatt Robert 
Published: January 2023

Abstract
Daclizumab, a monoclonal antibody that binds CD25, has been effective in the treatment of a number of autoimmune diseases. Despite continuous IL-2 receptor blockade for extended periods in these patients, there have been no instances of opportunistic infection and the rate of community-acquired infections is no greater than that of the general populace. This suggests that daclizumab dosing is balanced between prevention of autoimmunity and susceptibility to infection. To test this hypothesis, as a surrogate for infection we measured hepatitis vaccine responses in five patients with autoimmune uveitis who had been on daclizumab therapy for as long as 92 months. Surprisingly, only one equivocal vaccine response was observed. Given the high frequency of seroconversion (&gt;95%) induced by these vaccines in healthy adults, this study is sufficiently powered to demonstrate statistically significant differences from the expected hepatitis vaccine response rate (p=0.0005). These results show that daclizumab can block the response to a potent immunogen in vivo. Given that alum based vaccine responses are highly dependent on IL-4, the known inhibition of Th2 cytokine production by daclizumab is likely to have contributed to the vaccine failures we observed. Our results indicate that, where possible, antibiotic/antiviral drug prophylaxis may be preferable to vaccination in patients on daclizumab therapy.
Analysis of X-linked agammaglobulinemia patients in Mexico (B222)
Authors: Lopez-Herrera Gabriela , Berron-Ruiz Laura , Mogica Dolores , Espinosa-Rosales Francisco J , Santos-Argumedo Leopoldo 
Published: January 2023

Abstract
Humoral immunodeficiences are very common in the National Registry of Primary Immunodeficiences in Mexico. Among the humoral defects, X-linked agammaglobulinemia (XLA) is one of the most frequently found. In this work we report Btk mutations for 13 unrelated XLA patients some of wich has not been reported previously, most of the mutations affect Btk kinase domain, and PH and SH2 domain affecting mutations were also detected. Btk protein expression was found to be present in the majority of patients, in some cases we detect low expression levels and it was absent in other patients.