bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expression of KIT isoforms in systemic mastocytosis: correlation with disease severity and KITD816V (153.32)
Authors: Chan Ching , Bai Yun , Maric Irina , Klion Amy , Metcalfe Dean , Wilson Todd 
Published: January 2023

Abstract
Human KIT has several variants as the result of alternative mRNA splicing. Two major isoforms differ by the presence or absence of four amino acids (GNNK) at the juxta-membrane region of the extracellular domain. Somatic gain of function mutations in the tyrosine kinase domain of c-kit, most notably KITD816V, are detected in the majority of adult patients with systemic mastocytosis. We hypothesized that the expression pattern of GNNK variants may differ with disease severity, and transcripts containing the KITD816V mutation may segregate preferentially to one GNNK variant. A quantitative real-time PCR assay to assess GNNK- and GNNK+ transcripts was developed. We analyzed the ratio of GNNK- to GNNK+ transcripts in bone marrow mononuclear cells from 25 mastocytosis patients and 16 healthy controls. The GNNK- variant was more abundant in both patient and control samples. In advanced forms of mastocytosis, we observed a significant lower GNNK-/+ ratio compared to indolent systemic mastocytosis (p=0.0005). The KITD816V mutation was detected in both variants from KITD816V positive patients (n=21) and the KITD816V GNNK-/+ ratio was significantly higher than the total KIT GNNK-/+ ratio. Altogether, a relative increase in the GNNK+ variant was observed in advanced disease, and KITD816V specific transcripts displayed a relative GNNK- predominance. These data suggest that the determination of the GNNK-/+ ratio may be used to help determine disease prognosis.
Thymus-derived innate CD8+CD44hi T cells display features of antigen-experienced T cells with bystander helper properties (153.43)
Authors: Rafei Moutih , Williams Patrick , Ruiz Juan , Forner Kathy-ann , Dulube Gael , Galipeau Jacques , Perreault Claude 
Published: January 2023

Abstract
Innate CD8 T cells are abundant in several transcription factors (TFs)-deficient mouse models, but little is known about their development in wild-type animals. We used the RAG2-GFP mouse model to discriminate between memory, naïve classic and innate TCRαβ CD8 cells in the thymus. Relative to classic T cells, innate thymocytes display increased levels of numerous cell surface molecules (TCR, CD2, CD3, CD5, CD38, CD62L, B7H1) and enhanced expression of the TFs T-bet, EOMES, Id3 and KFL2. Hematopoietic chimera studies revealed that non-hematopoietic MHCI+ cells are required for innate CD8 T cell selection in the thymus. Mice expressing the transgenic OTI TCR generate both classic and innate CD8 thymocytes suggesting that both subsets have overlapping TCR repertoires. Innate CD8 T cells were also found to rapidly produce high levels of IFNγ upon stimulation and to readily proliferate in response to IL2 and IL4. Under lymphopenic conditions, classic CD8 T cell expansion was 2-3 fold greater than that of innate T cells, which constitutively express B7H1 making them unduly susceptible to apoptosis. To test their bystander properties, we added classic or innate T cells as third-party cells to an in vitro antigen presentation assay. Innate T cells were capable of promoting the proliferation and activation of responding T cells in a CD44-dependent manner. These observations show that innate CD8 T cells represent a subset of thymocytes that differ in many ways from classic CD8 T cells.
CD28 costimulation is not required for the development and differentiation of γδ T cell effectors (153.45)
Authors: Laird Renee , Wolf Benjamin , Princiotta Michael , Hayes Sandra 
Published: January 2023

Abstract
Both antigen recognition and CD28 costimulation are required for the activation of naïve αβ T cells and their subsequent differentiation into cytokine-producing or cytotoxic effectors. However, because of contradictory results, it remains unresolved as to whether CD28 costimulation is also necessary for the activation and differentiation of γδ T cell effectors. Recently, it has been shown that γδ T cells acquire effector functions in the thymus following encounter with self-ligands. This is in contrast to αβ T cells, which acquire effector functions in secondary lymphoid tissues following encounter with foreign antigens. In light of these newly identified requirements for γδ effector cell development, we re-examined the role of CD28 in γδ T cell effector fate specification and function. We report that, although IFNγ-producing γδ effectors (γδ-IFNγ) express higher levels of CD28 than IL-17-producing γδ effectors (γδ-17), CD28-deficiency has no effect on the development of either γδ-17 or γδ-IFNγ cells. Also, following infection with Listeria monocytogenes, we found that the activation, expansion and differentiation of γδ-17 and γδ-IFNγ cells were normal in CD28-/- mice. Thus, these data not only indicate that CD28 is not essential for γδ T cell effector development and differentiation but also highlight differences in the requirements for the generation of αβ and γδ T cell effectors.
NFIL3/E4BP4 is a key transcription factor for CD8α+ dendritic cell development (153.47)
Authors: Kashiwada Masaki , Pham Nhat-Long , Pewe Lecia , Harty John , Rothman Paul 
Published: January 2023

Abstract
Antigen presentation by mature dendritic cells (DCs) is the first step for initiating adaptive immune responses. DCs are comprised of heterogeneous functional subsets. However, the molecular mechanisms regulating differentiation of specific DC subsets are not understood. Here, we report that the basic leucine zipper transcription factor NFIL3/E4BP4 is essential for the development of CD8α+ conventional DC (cDC). Nfil3-/- mice specifically lack CD8α+ cDCs but not CD8α- cDCs or plasmacytoid DCs in lymphoid tissues. Flt3 ligand-dependent generation of CD8α+ cDCs in lymphoid tissues and CD8α+-equivalent cDCs from Nfil3-/- bone marrow cells was also impaired. NFIL3 regulates CD8α+ cDC development in part throuth Batf3 expression. Importantly, Nfil3-/- mice exhibit impaired cross-priming of CD8+ T cells against cell-associated antigen, a process normally carried out by CD8α+ cDCs, and failed to produce IL-12 after TLR3 stimulation. Thus, NFIL3 plays an essential role in the development of CD8α+ cDCs.
Human thrombopoietin knockin mice efficiently support human hematopoiesis in vivo (153.5)
Authors: Rongvaux Anthony , Willinger Tim , Takizawa Hitoshi , Rathinam Chozhavendan , Eynon Elizabeth , Stevens Sean , Manz Markus , Flavell Richard 
Published: January 2023

Abstract
Hematopoietic stem cells (HSCs) both self-renew and give rise to all blood cells for the lifetime of an individual. Xenogeneic mouse models are broadly used to study human hematopoietic stem and progenitor cell biology in vivo. However, maintenance, differentiation, and function of human hematopoietic cells is suboptimal in these hosts. We hypothesized that this defect was due to reduced cross-reactivity of mouse cytokines on the human receptor. Thrombopoietin (TPO) has been demonstrated as a crucial cytokine supporting maintenance and self-renewal of HSCs. We generated RAG2-/-γc-/- mice in which we replaced the gene encoding mouse TPO by its human homologue, and we transplanted human CD34+ cells in these recipients. Homozygous humanization of TPO (TPOh/h) led to increased levels (higher than 80%) and longer maintenance (more than 6 months) of human cell engraftment in the bone marrow of the hosts. Multilineage differentiation of hematopoietic cells was also improved, with an increased ratio of myelomonocytic verus lymphoid lineages. Moreover, maintenance of human stem and progenitor cells was improved : we observed a significant increase in the fraction of human Lin-CD34+CD38-CD90+CD45RA- cells in TPOh/h compared to control recipients; and CD34+ cells purified from TPOh/h primary recipient had a significantly increased capacity to efficiently engraft secondary recipients. Therefore, RAG2-/-γc-/- TPO-humanized mice represent a novel model to study human hematopoiesis in vivo.
Increased lethality but decreased encephalitis in IRF8-deficient mice following West Nile Virus infection (154.1)
Authors: Minten Carsten , Terry Rachael , King Nicholas , Campbell Iain 
Published: January 2023

Abstract
The transcription factor IRF8 is crucial in myeloid and dendritic cell development and the response of these cells to the cytokine IFN-γ. The absence of IRF8 is associated with specific morphological and functional changes in peripheral macrophages as well as CNS resident myeloid cells known as microglia. These cells have been shown to be pivotal in the pathogenesis of West Nile Virus (WNV) encephalitis in mice. Therefore, we examined the role of IRF8 in the host response to infection of the brain with WNV. IRF8 KO mice showed increased lethality following intranasal challenge with low dose WNV compared with similarly infected WT mice. In the brain, WNV loads were higher and virus more disseminated, however, there was a dramatic reduction of infiltrating T-cells and macrophages in WNV-infected IRF8 KO mice compared with similarly infected WT animals. While the cerebral expression of many proinflammatory chemokine (CCL2, CCL4, CCL5, CXCL10) and cytokine (TNF and IL-6) genes was significantly increased in both WT and IRF8 KO mice infected with WNV, expression of the IL-12p40 and IFN-γ genes was increased in WT but not IRF8 KO mice. In conclusion, IRF8 is apparently dispensable in the localised innate inflammatory response to WNV infection, however, IRF8 has a key role in the antiviral response involved in controlling virus replication and spread within the CNS. Furthermore, our data demonstrates that IRF8 may be crucial for the adaptive immune response against WNV in the CNS.
Cationic Lipid DNA Complex (JVRS-100) treatment of woodchucks with Hepatitis B virus induced Hepatocellular Carcinoma (154.12)
Authors: Fairman Jeffery , Liu Katherine , Menne Stephan 
Published: January 2023

Abstract
To determine the dependence of responsiveness to immune stimulation on the woodchuck hepatitis B viral (wHBV) load, we evaluated cytokine and cell surface marker mRNA expression following dosing of JVRS-100 in chronic woodchuck carriers with low (mean: 2.2x1010 ge/ml), medium (4.0x1010 ge/ml) and high (4.2x1011 ge/ml) serum viral loads. Our hypothesis was that high viral loads seen in chronic wHBV infection (10x human) result in immune suppression to external immune stimuli. Woodchucks were dosed intravenously with JVRS-100 and evaluated for immune activation. Woodchucks with low viral load had increased expression of cytokine and cell surface markers in contrast to animals with moderate or high viral loads with no change in expression. To evaluate anti-tumor effects of JVRS-100 we dosed groups of woodchucks with 100µg or 300µg JVRS-100. When normalized for pre-treatment levels of wHBV serum DNA, there was a steady decrease in viral load of treated woodchucks. There was no difference in the tumor burden; however, woodchucks in the high dose JVRS-100 group developed no new tumors over the 24 weeks in which they were monitored (12 weeks Rx followed by 12 weeks observation). This is unusual for this disease model and we believe highly significant to the development of an intervention for HBV induced HCC. Our working hypothesis is that the administration of JVRS-100 blocks the conversion of chronic HBV infection to HCC.
Reduction of HIV-1 innate immune activation by protease inhibitors (154.15)
Authors: Wallet Mark , Appelberg Sofia , Reist Caroline , Guiulfo Giorgio , Sleasman John , Goodenow Maureen 
Published: January 2023

Abstract
HIV-1-associated innate immune activation is caused by viral replication and/or microbial substances that leak from the gut lumen into the circulation (microbial translocation). Consequences of the ensuing HIV-1-associated inflammation range from neurocognitive impairment to cardiovascular disease where macrophages play a central role. HIV-1 therapy is evaluated based on suppression of viremia and restoration of CD4 T cells, while the effects of therapy on innate immune activation are rarely considered. The current study assessed the effects of protease inhibitors on macrophage activation ex vivo and in vivo. Nelfinavir [NFV] and tipranavir [TPV] inhibited LPS activation of human macrophages ex vivo as measured by reduced secretion of LPS-binding protein soluble CD14 [sCD14] and pro-inflammatory cytokines IL-6 and TNF. Cytokine secretion was inhibited prior to transcription by modulation of toll-like receptor signaling pathways. In vivo, NFV-treated HIV-1-infected patients presented with significantly reduced monocyte/macrophage activation as measured by a greater reduction in sCD14 plasma levels compared to those treated with indinavir or ritonavir. These data suggest that anti-inflammatory effects of NFV or TPV may augment efficacy of HIV-1 treatment by targeting HIV-1-assocated innate immune activation. Delineation of cellular targets of NFV or TPV will lead to novel approaches to suppress innate immune activation in HIV-1, other infectious diseases or autoimmunity.
Epstein-Barr Virus LMP2A modulates B cell proliferation and cytokine production (154.16)
Authors: Swanson-Mungerson Michelle , Incrocci Ryan , Bultema Rebecca 
Published: January 2023

Abstract
Despite the association of Epstein-Barr Virus (EBV) with numerous malignancies and autoimmune diseases, the mechanism by which EBV promotes disease is unknown. After initial infection, EBV transitions to a latent state in which few viral transcripts, such as Latent Membrane Protetein 2A (LMP2A), are expressed. While LMP2A is known to promote B cell survival, our data using primary LMP2A-expressing B cells indicate that LMP2A increases B cell proliferation in response to mitogens. Since cytokines are critical in promoting B cell survival and proliferation, we hypothesized that LMP2A increases B cell cytokine production to increase their survival and proliferation. Our data indicate that LMP2A does not globally increase B cell-produced cytokine levels, but rather targets a limited number of cytokines. These findings implicate that LMP2A-expressing B cells not only have the ability to influence B cell survival and proliferation in an autocrine manner, but may also secrete cytokines to modulate immune cells in their environment. This work is funded by NIH grant 1 R15 CA149690-01 and Midwestern University Intramural Funds.
Glycan-induced antiviral immune response inhibits HIV replication (154.19)
Authors: Ring Jennifer , Harn Don 
Published: January 2023

Abstract
Several viruses have developed strategies to evade recognition by the host immune response, thereby establishing chronic infection. In these cases, an immunotherapeutic drug that triggers an antiviral immune response could produce a more favorable outcome. We evaluated the immunomodulatory human milk glycan Lacto-N-fucopentaose III (LNFPIII) for its ability to trigger an antiviral immune response and inhibit HIV replication. Human PBMCs were treated with LNFPIII-HSA, a glycoprotein consisting of on average, 8-10 molecules of LNFPIII conjugated per molecule of HSA via an APD linker. This treatment resulted in the upregulation of several antiviral cytokines including TNF-α, IFN-γ, and major HIV-suppressive chemokines MIP-1α, MIP-1β, and RANTES. Upon infection, we observed reduced HIV replication in treated PBMCs, likely due to the multitude of antiviral cytokines present. When monocytes were removed from culture, or CD14 was neutralized, the antiviral effects of LNFPIII-HSA were negated. These results indicate that LNFPIII-HSA triggers an antiviral immune response capable of controlling HIV replication by activating the antiviral activity of CD14+ monocytes.
Human cytomegalovirus decreases constitutive transcription of MHC class II genes in mature Langerhans cells by reducing CIITA transcript levels (154.32)
Authors: Lee Andrew , Hornell Tara , Wang Nan , Harding James , Deshpande Chetan , Hertel Laura , Lacaille Vashti , Pashine Achal , Macaubas Claudia , Mocarski Edward , Mellins Elizabeth 
Published: January 2023

Abstract
Human cytomegalovirus (HCMV) productively infects CD34+ progenitor-derived, mature Langerhans-type dendritic cells (matLC) and reduces surface expression of MHC class II complexes (MHC II) by increasing intracellular retention of these molecules. To determine whether HCMV inhibits MHC II expression by other mechanisms, we assessed mRNA levels of the class II transcriptional regulator, CIITA, and several of its target genes in infected matLC. CIITA mRNA levels were significantly lower in HCMV-infected matLC than in mock-infected cells. Analysis of promoter-specific CIITA isoforms revealed that types I, III and IV were decreased by HCMV, a result not seen after incubation of matLC with LPS. Exposure to UV-inactivated virus failed to reduce CIITA mRNA levels, implicating de novo viral gene expression in this effect. The stability of CIITA mRNA was not diminished by HCMV, arguing that diminished transcription is the mechanism for CIITA mRNA reduction. HCMV-infected matLC expressed lower levels of DR transcripts and reduced DR protein synthesis rates compared to mock-infected matLC. Thus, we demonstrate that HCMV infection of a human dendritic cell subset inhibits constitutive CIITA expression, resulting in reduced MHC II biosynthesis as a candidate immunoevasive mechanism.
Abrogation of human CD59 function regains neutralizing and non-neutralizing antibody functions in triggering complement-mediated virolysis of HIV-1 (154.7)
Authors: Yu Qigui , Hu Ningjie , Amet Tohti , Byrd Daniel 
Published: January 2023

Abstract
Patients infected with HIV-1 are well known to mount a vigorous and sustained antibody (Ab) response to the virus, yet fail to control virus proliferation or protect them from developing AIDS. Recent studies have suggested that resistance of HIV-1-infected cells and virions to Ab-dependent complement-mediated lysis (ADCML) is dependent on regulators of complement activation (RCA), particularly CD59 that controls the formation of the membrane attack complex (MAC) at the terminal stage of the complement activation cascades via all three activation pathways. Abrogation of CD59 function renders HIV-1 virions and infected cells sensitive to ADCML. In the current study, we used the well-characterized anti-HIV-1 nAbs (2G12, 2F5, 4E10 and b12) and non-nAbs (5F3, A32 and F240) to investigate whether the enhancement of ADCML by inhibition of CD59 is mediated by nAbs, non-nAbs or both. We found that these Abs, regardless of their neutralizing activity, significantly enhanced ADCML of many HIV-1 lab strains and 6 primary isolates examined in the presence of CD59 blockers. Direct addition of CD59 blockers into plasma from 16 HIV-1-infected individuals triggered ADCML of endogenous plasma virions, and the effect correlated to Ab titers rather than Ab neutralizing activity, indicating that patient plasma Abs are able to recognize endogenous virions, and abrogation of CD59 may represent a novel approach to regain the function of both nAbs and non-nAbs naturally present in patient’s blood.
Identification and characterization of CD8+ T helper cells based on CD40L expression (155.1)
Authors: Frentsch Marco , Stark Regina , Listopad Joanna , Meier Sarah , Schulz Axel , Durlanik Sibel , Blankenstein Thomas , Thiel Andreas 
Published: January 2023

Abstract
CD8+ T cells are primarily regarded as cytotoxic cells. We have identified a substantial subset of CD40L expressing non-cytotoxic T-cells among primary human CD8+ T cells. CD40L+ CD8+ T cells exert diverse characteristic Th-cell functions such as activation of B cells, induction of DC maturation and secretion of cytokines such as IL-2, IFNγ, TNFα or IL-4. Up to 25% of total human central and effector memory CD8+ T cells express CD40L including cells specific for pathogens such as influenza, CMV and EBV, as well as yellow fever virus specific cells after primary vaccination. At next we assessed the functional relevance of CD40L expression on CD8+ T cells during a anti-tumor response. We challenged Rag1-/- mice with SV40 Tag expressing tumor cells and injected in parallel wt or CD40L-/- CD8+ T cells. Only application of CD40L competent wt CD8+ T cells prevented the establishment of tumors, whereas injection of CD40L-/- CD8+ T cells resulted in non-controlled tumor progression similar to non-treated tumors, although tumor-specific T cells were primed in these mice. Our results disclose an essential functional relevance of CD40L expressed by CD8+ T cells. Especially, in situations of reduced CD4+ T-cell help and MHC-II antigen presentation and/or limited danger signals CD40L+ CD8+ T cells may exert essential helper responsibilities for immunity and thus are potent candidate T cells to execute or support effective anti-tumor or anti-pathogen immune therapies.
Immune adjuvant and antitumor activity of baccatin III, a synthetic precursor of taxol (155.15)
Authors: Kim Ki-Hyang , Park Chan-Su , Park Dong-Eun , Im Sun-A , Lee Chong-Kil 
Published: January 2023

Abstract
Baccatin III (BA-III), a precursor for the synthesis of taxol, is widely considered to be an inactive derivative of taxol. Here we show that baccatin III exerts immune adjuvant activity. When added to cultures of immature dendritic cells (DCs) generated from bone marrow cells with GM-CSF, baccatin III increased the expression of MHC II molecules and the allostimulatory activity of the DC. The immune adjuvant activity of baccatin III was examined in mice injected with a model antigen, OVA, emulsified with Freud’s incomplete adjuvant. Inclusion of baccatin III significantly increased the production of OVA-specific IgG, but not IgM. The in vivo combination effects of baccatin III and gemcitabine, an antitumor agent, were examined in mice bearing mammary carcinoma 4T1 cells. Combination of baccatin III with gemcitabin efficiently inhibited the tumor growth and the accumulation of myeloid derived suppressor cells (MDSC, CD11b+Gr-1+ cells), while increasing cellularity of CD4 T cells and CD8 T cells in the spleen, compared to baccaatin III only- or gemcitabine only-treated groups. These results demonstrate that baccatin III exerts immune adjuvant activity, and further show that baccatin III in combination with gemcitabine, a nucleoside analog used as chemotherapy, may be useful in the treatment of tumors that induce the accumulation of MDSCs.
Computer analysis and identification of antigenic determinants from five glycoproteins of HSV2 (155.17)
Authors: Yuexi Li , Xingsheng Wang , Jianmin Liao , Dengke Yin , Suqin Li , Zhengmao Wang , Lin Li , Guangyan Xie 
Published: January 2023

Abstract
To screen the antigenic determinants from glycoprotein B (gB2), C (gC2), E (gE2), G (gG2) and I (gI2) of herpes simplex virus II (HSV2) for designing novel vaacine, the aminao acid sequences of the five proteins were analysed with several software algorithms, DNAstar, Biosun, and Antheprot. The common antigenic determinants screened by all the software algorithms were synthesized chenically and identified by animal immune experiments. Sixteen synthesized antigenic determinants selected from the five proteins were linked to carrier protein BSA respectively to enhance their immunogenicity, and uesed as immunogens to immunize mice. The antibodies in the sera of the immunized mice were detected with the coreesponding immunogens and the parent glycoproteins by EIA and western blotting. The virus-neutralizing activity of the antisera were tested with vero cell in vitro. The antisera reacted strongly with the parent glycoproteins, and also the peptides reacted strongly with antibodies against the parent glycoproteins. The antisera against five antigenic determinants from gB2 ,gC2, gE2, gG2, and gI2 can neutralize HSV2 infectivity in vitro respectively, which has not been reported up to now. These results suggested that the antigenic determinants from the five glycoproteins screened by software algorithms and validated by experiments are promising for vaccine design.