bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human natural killer cell expansion in vitro using mitomycin-C treatment as an alternative to irradiation of modified K562 feeder cells.
Authors: Carr William H , Zinsou Carmen , Rowe Tahishia , Lange Christopher , Lawal Oluwadamilola O. 
Published: January 2023

Abstract

Introduction
Current methods for in vitro expansion of Natural Killer (NK) cells require feeder cell irradiation, which is not readily available in many biomedical research or clinical facilities. Here we evaluated mitomycin-C (MMC) treatment of feeder cells as an alternative to irradiation for the purpose of expanding NK cells in human peripheral blood mononuclear cell (PBMC) cultures.

Methods
Using PBMCs from 4 healthy blood donors and genetically modified K562 feeder cells (K562-mb21-41BBL cells) that preferentially stimulate NK cells, we evaluated the efficacy of MMC treatment in limiting the proliferation of the feeder cells at a range of doses (0 ug/mL, 2 ug/mL, 10 ug/mL, 20 ug/mL, 30 ug/mL, and 200 ug/mL) with 3-hour incubation. We used 7AAD dye and CFSE proliferation dye to measure viability and cell proliferation, respectively. We then compared the percentages of NK cells (CD3neg, CD56pos) in PBMCs cultured for 14 days in the presence of MMC-treated feeder cells or irradiated feeder cells using multiparametric flow cytometry.

Results
We found that 10 ug/mL dose of MMC for 3-hrs achieved a significantly higher percentage of viable cells compared to higher doses (p&lt;0.05, ANOVA), and also significantly limited proliferation compared to untreated feeder cells. Furthermore we found no significant difference in NK cell expansion by 10 ug/mL MMC treated feeder cells and 100 grays irradiated feeder cells.

Discussion
In conclusion, we found that 10 ug/mL mitomycin-C treatment of modified K562 feeder cells was optimal for limiting their proliferation for NK cell expansion that was equivalent to using irradiated feeder cells.
Identification of kidney CD45intCD11bintF4/80+MHCII+Ly6C− macrophages initially masquerading as T cells
Authors: Lee Sul A , Noel Sanjeev , Sadasivam Mohanraj , Hamad Abdel Rahim , Rabb Hamid 
Published: January 2023

Abstract

Background
Mononuclear phagocytic cells (MPCs) have important roles in the pathogenesis of many kidney diseases, including ischemia-reperfusion injury (IRI) and allograft rejection. Despite the recent advances on description of MPC subpopulations and their functional characterization in kidney, there is no clear consensus for the classification of MPC subpopulations. Here we describe a newly identified renal macrophage, named CD45intCD11bint cells.

Methods
Kidney mononuclear cells were isolated from C57BL6 male mice under steady state and after IRI and analyzed using flow cytometry.

Results
While focusing on TCRαβ+CD4−CD8− kidney T cells, we identified a cell population that binds only to TCRβ and CD8β antibodies but not to CD8α antibody. Further studies using Fc receptor blockers disclosed that this population was a renal macrophage subset which was different from other macrophages by its intermediate expression of CD45 and CD11b. These CD45intCD11bint macrophages were further characterized as F4/80+MHCII+Ly6C− cells comprising 40% of MPCs in normal kidney. CD45intCD11bint macrophages are found predominantly in the kidney as compared to other lymphoid and non-lymphoid organs (ANOVA, p = 0.002). In addition, CD45intCD11bint population significantly decreased 48 hours after IRI in contrast to steady increase of CD45high macrophages (p = 0.027). Relevance of this population to human kidney is being investigated.

Conclusion
Kidney CD45intCD11bintF4/80+MHCII+Ly6C− macrophages are newly defined MPCs that comprise a major subset of resident renal MPCs. They have unique surface phenotype as well as T-cell mimicking characteristics. This new subset could play an important role in immune diseases of the kidney.
Regulatory B cells in the blood of plasma cell hepatitis patients who have undergone liver transplantation
Authors: Hasan Md Mahmudul , Joo HyeMee , Klintmalm Goran B. , Oh SangKon , Thompson-Snipes LuAnn 
Published: January 2023

Abstract
Alloreactive T cells and antibody-mediated rejection (AMR) appear to be barriers to long-term organ engraftment. Regulatory B cells (Bregs), a B cell subset, can protect against autoimmune/inflammatory damage, in part, by producing IL-10, an anti-inflammatory cytokine. However, the role of Bregs in solid organ transplantation is poorly understood. In this study, we examined IL-10 producing human Bregs in the blood of healthy controls compared to liver transplanted patients with plasma cell hepatitis (PCH), a variant of late onset rejection. First, we optimized conditions for human Breg differentiation in vitro. Based on the presence of differentially expressed cell surface markers on IL-10+ CD19+ B cells, we then established a profile of Bregs from several human tissues including tonsils and spleens and demonstrated that in vitro generated IL-10+CD19+ B cells are capable to suppress alloantigen stimulated T cell proliferation and cytokine production. Finally, we find that the frequency of CpG B activated IL-10+CD19+ B cells is negligible in PCH+ liver transplant patients compared with healthy controls. In line with this, the frequency of transitional B cells (CD19+CD24hiCD38hi) is significantly lower in PCH+ liver transplanted patients than healthy controls. Our findings suggest that a lack of immunosuppressive Bregs could be associated with the rejection in the PCH+ patients
The Intracutaneous Absorption of Antigen
Authors: Smith G. H. , Cook M. W. 
Published: January 2023

Abstract
That the body is capable of reacting to parenterally introduced antigenic substances has been amply proven, but as yet there has been no satisfactory explanation of the mechanism of the interaction, that is, it has not been conclusively determined whether antigen is eliminated as antigen or whether it enters into the metabolic processes and becomes utilized or denatured. It can not be assumed that identical processes follow the introduction of different antigens or even of the same antigen when injected by different routes. In fact, opinion differs very widely as to what happens when a given antigen is introduced by a single route. Further complications arise through the fact that animals receiving antigen may, as a result of inherent peculiarities or experimental procedure, be endowed with a variety of metabolic forces, which in individual cases may so combine as to give a variety of results.
Experiments with a Possible Bearing Upon the So-Called Non-Specific, Intravenous Vaccine Therapy
Authors: Teague Oscar , McWilliams Helen I. 
Published: January 2023

Abstract
The experimental foundation for a non-specific vaccine therapy was laid by Emmerich (1) thirty years ago. He showed that rabbits inoculated with streptococci possessed a certain amount of immunity against anthrax. He also observed that an anthrax infection in rabbits, even after it had called forth distinct symptoms, could be favorably influenced by the injection of streptococci—particularly if the latter injection was administered intravenously. Pawlovsky (2) confirmed these results, and demonstrated that Friedländer's bacillus and B. prodigiosus also retarded the development of the anthrax infection. Bouchard (3) found that B. pyocyaneus could be used for this purpose. Then Freudenreich (4) and Woodhead and Cartwright Wood (5) obtained similar results with killed cultures of B. pyocyaneus, and Buchner (6), with killed cultures of Friedländer's bacillus. Von Dungern (7) injected killed cultures of Friedländer's bacillus intravenously and observed that rabbits so treated were more resistant to anthrax bacilli inoculated subcutaneously one or several days later, than were normal rabbits.
The Differentiation of a Type of Human Blood by Means of Normal Animal Serum
Authors: Landsteiner K. , Levine Philip 
Published: January 2023

Summary
The main result of the present investigation consists in making available a method—namely, the use of normal animal serum—for readily differentiating a certain rather frequent type of human blood designated as P, whereas the reagents previously employed for this purpose were not easily accessible.
The method may be of use for studies on heredity and racial distribution.
The Antigenic Properties of Fibrinogens
Authors: Kyes Preston , Porter Robert T. 
Published: January 2023

Abstract
Previous publications (1) (2) from this laboratory have pointed out the inadequacy of the usual methods of blood defibrination in preparing fibrinogen-free serum and particular methods have been given by which such a serum may be obtained. The fact has also been established that traces of fibrinogen in a serum profoundly modify the immune reactions of that serum both in vivo and in vitro, provoking shock reactions in the former and pseudoprecipitin and agglutinin reactions in the latter.
Continuing the study of fibrinogen in relation to immune reactions we have recently tested the antigenic properties of fibrinogen and this paper presents evidence that the injection of foreign fibrinogen into an animal induces the production of antibodies specific for the particular fibrinogen used.
The fibrinogen chiefly employed in our experiments was prepared from well developed Plymouth Rock cockerels.
Comparative Study of the Anaphylactic and Tuberculin Types of Hypersensitiveness
Authors: Dienes L. 
Published: January 2023

Summary
The crystalline egg albumin and egg globulin differ considerably in their relationship to the different manifestations of hypersensitiveness. The crystalline egg albumin tends to sensitize the organism to those manifestations of the hypersensitiveness which are mediated by antibodies. The egg globulin tends to sensitize to those in which the active immunization plays the more important rôle. The difference between the antigens lies in their capacity to produce a different type of sensitization. We find no marked difference between them concerning their capacity to produce antibodies or to elicit the reactions themselves. The observations with crystalline egg albumin and egg globulins present an analogy to the behavior of certain bacterial antigens which we usually find connected either with the tuberculin or with the anaphylactic type of sensitiveness.
We have given a short review of our experiences concerning the capacity of different antigens to produce the tuberculin type of skin sensitiveness. With all innocuous antigens which we tested—as eggwhite and animal sera—we succeeded in producing this sensitiveness. With bacterial antigens we have not so far succeeded in producing it.
Over-expression of BDNF in Primary Mouse B cells
Authors: Torres Vanessa , Matsui Takeshi K. , Stowe Ann Marie 
Published: January 2023

Abstract

Background
Neuronal networks require significant neurotrophic support for recovery after stroke, but delivery of neurotrophins (NTs) failed in clinical trials due to short half-lives and limited brain permeability. B cells, however, infiltrate the post-stroke brain and possess the ability to produce NTs (e.g. brain-derived neurotrophic factor (BDNF)). Thus, we hypothesized that neurotrophic B cells could enhance neuroplasticity after stroke.

Methods
A CMV-BDNF-mCherry plasmid was transfected into HEK-293T cells or LPS-stimulated splenic B cells isolated from 6–10 week old C57Bl/6J mice. Transfected B cells were co-cultured with supporting splenocytes for 24, 48, or 72h. Flow cytometry assessed mCherry+ B cells and BDNF secretion was measured via ELISA. Graphpad Prism analyzed statistical differences. Significance was p&lt;0.05.

Results
The CMV-BDNF-mCherry plasmid was successfully transfected into HEK-293T cells compared to non-transfected controls, and led to long-term BDNF secretion (48h-3,162.9pg/mL, 72h-5,106.7pg/mL; both p&lt;0.05 vs. 24h-720pg/mL). For splenic B cells, the percentage of transfected mCherry+ B cells was highest at 24h after transfection (12.6% of CD45+CD19+ cells) and reduced by 48h (8.77%; p&lt;0.01 vs. 24h) and 72h (6.03%; p&lt;0.01 vs. 24h). This trend was also reflected in absolute mCherry+ B cell numbers. Despite this reduction in mCherry+ B cells, BDNF secretion was consistently detected over 72h (24h-303.3pg/mL, 48h-453pg/mL, 72h-317.6pg/mL).

Conclusions
Our studies show successful overexpression of BDNF in primary B cells. We are currently assessing the magnitude of B cell-derived neurotrophic support on neuroplasticity under homeostatic conditions and in vitro stroke injury models.
Humoral Immune Response Modulation by Aryl hydrocarbon Receptor Activation (AhR) in CD11c+ Cells: Exploring Gender-specific and Ligand-dependent Effects on Antibody Responses in Mice.
Authors: Kreitinger Joanna , Shepherd Celine Ann Beamer , Shepherd David M. 
Published: January 2023

Abstract
Modulation of antibody responses is often a summation of effects on B cells, helper T cells, and dendritic cells (DCs). All three of these immune cells express AhR, a ligand-activated transcription factor with endogenous and exogenous ligands known to modulate immune responses. Importantly, antibody responses are keenly sensitive to AhR activation by TCDD, its prototypical ligand and common environmental pollutant. However, the specific contribution of AhR activation in DCs has not yet been defined, particularly in the context of gender-specific and ligand-dependent effects. We aimed to determine the effects of AhR activation in DCs by endogenous and exogenous ligands on OVA-specific antibody responses in mice. Specifically, we hypothesized that AhR activation in DCs alone suppresses antigen-specific serum antibody production, selectively affecting different isotypes. To test this hypothesis, we used mice conditionally deficient of AhR in CD11c+ DCs (AHRCD11c). Control (AhRd) and AHRCD11c mice were treated with vehicle, ITE or TCDD via oral gavage, and subsequently immunized i.p. with whole OVA/adjuvant to induce a systemic, antigen-specific humoral response. One week later, AhRd mice showed significantly reduced OVA-specific IgM and IgG2a titers while IgA was significantly increased; effects that were ameliorated in AhRCD11c mice. Interestingly, TCDD-induced IgA increases were both AhR- and gender-dependent with female mice showing increased IgA production. Collectively, these results demonstrate that DCs contribute to, but are not wholly responsible for, AhR-mediated effects on antigen-specific serum antibody generation. Research supported by ES013784 (DMS) and AAI Careers in Immunology Fellowship (JMK).
Changes in metabolic phenotype and cellular ATP production during CD4+ T cell activation.
Authors: Romero Natalia , Swain Pamela , Kam Yoonseok , Dranka Brian P. 
Published: January 2023

Abstract
Activation of CD4+ T cells is followed by rapid proliferation and differentiation into specific subsets (Treg or Teff). These transitions are accompanied by tight regulated changes in energetic demand and cellular metabolic reprogramming. Using Agilent Seahorse extracellular flux analysis, we have developed a cell-based assay for simultaneous measurement of the two-main cellular ATP-producing pathways, i.e. glycolysis and oxidative phosphorylation. The assay allows for quantification of real time changes in total ATP production rate, and the fractional contribution of the individual pathways to support bioenergetic demands. We found that naïve CD4+ T cells obtain most of its ATP from mitochondrial oxidative phosphorylation. After activation with anti CD3/CD28 conjugated beads, total ATP production rate significantly increases, and that rise is sustained by an increase in glycolytic ATP production, but also in mitochondrial ATP production. When glucose is replaced by galactose in the assay medium, T cell activation is inhibited despite partial compensation of total ATP production through mitochondrial respiration. Time-course of bioenergetic phenotype during T-cell expansion shows a deep increase in ATP production rate up to day 6–7 with glycolysis contributing up to 70% of total ATP production. When glucose is replaced by galactose 72 hr after activation, total ATP production rate was fully compensated by mitochondrial ATP production but still the inhibition of glycolytic activity by galactose reduces cell proliferation. The results obtained highlight the role of glycolytic activity beyond energetic supply and its importance for biosynthetic and redox requirements or cell activation and proliferation.
Physiological microbial exposure has the capacity to substantially influence inflammatory responses in vivo
Authors: Sjaastad Frances V. , Pierson Mark , Huggins Matthew A. , Danahy Derek B. , Badovinac Vladimir P. , Griffith Thomas S. , Hamilton Sara E. 
Published: January 2023

Abstract
Physiological exposure to microbes profoundly effects the basal immune state, based on data from ‘dirty’ mice with a diverse microbial experience obtained after co-housing with pet store mice. Most T cells in dirty mice and adult humans have an effector/memory phenotype, but specific pathogen free (SPF) mice or newborn humans have few Ag-experienced memory T cells. These differences prompted us to define the acute cytokine/chemokine response in SPF or co-housed inbred C57Bl/6 (B6) mice. Co-housing for 60 d increased the frequency of CD44hi CD8 T cells and led to higher basal serum cytokine/chemokine levels (vs. SPF mice). We then examined to what extent these basal immune differences affected the response to different inflammatory stimuli. Infection of SPF mice with virulent Listeria significantly increased serum cytokines/chemokines (e.g., IFNg, TNF, CXCL1, CXCL10, CCL7) 24–72 h after infection. The same infection produced little-to-no change in the serum of dirty mice, but they had significantly lower bacterial burden. After LPS challenge, SPF and dirty mice had similar increases in serum cytokines/chemokines, but mortality was increased in dirty mice. Lastly, while an increase in serum cytokines/chemokines occurred 6–12 h after sepsis induction (CLP model) in both SPF and dirty mice, they were 2–8-fold higher in dirty mice. Acute lymphopenia was observed in both sets of CLP-treated mice; yet, mortality was higher in CLP-treated dirty mice. Thus, physiological microbial exposure influences the cytokine ‘storm’ and survival suggesting the immune status of the host controls the magnitude of the response and outcome. The data also point out the need to further develop the ‘dirty’ mouse model to complement current studies in SPF mice.
CD28 co-stimulation drives memory phenotype (MP) Treg cell and MP CD4+Foxp3− effector T cell homeostatic proliferation.
Authors: Panda Abir Kumar , Shevach Ethan M. 
Published: January 2023

Abstract
Foxp3+Tregs, CD4+Foxp3− effector T cells and CD8+ T cells are composed of naïve (NP, CD44loCD62Lhi) and memory (MP, CD44hiCD62Llo) subsets. About 30% of MP T cells are cycling (Ki-67+) in-vivo. To determine the factors that drive T cell homeostatic proliferation in-vivo, we investigated the roles of cytokines (TNFa, IL-2, IL-7, and IL-33), TCR-MHC-II interactions, co-stimulatory (CD28, ICOS, CD40, GITR, OX40), and co-inhibitory (CTLA-4, PD1, BTLA, TIGIT) receptors. Blockade of CD28-CD80/86 signaling completely inhibited MP Treg and MP CD4+Foxp3− effector T cell proliferation but did not affect MP CD8+ T cell proliferation. Inhibition of cytokine activity did not affect the proliferation of any of the T cell populations. Marked enhancement of homeostatic proliferation of MP Treg and MP CD4+Foxp3−T cells was observed by blocking TCR-MHCII interactions, CTLA4-CD80/CD86 interactions, or by agonistic mAbs to CD40, GITR, and OX40. Blocking PD1-PDL1 interactions modestly enhanced proliferation, while blocking BTLA, TIM3, ICOS and TIGIT did not affect MP T cell proliferation. Importantly, blocking of CD28-CD80/86 signaling completely abrogated the augmentation of proliferation observed after blocking TCR and CTLA4, PD1, or by agonistic mAbs to GITR, OX40, and CD40. Our results support a complex pathway in which CD28-CD80/CD86 interactions play a critical role in Treg and MP CD4+Foxp3−T cells proliferation in-vivo. CTLA4-CD80/CD86 interactions act as a brake, while MHCII-TCR interactions negatively regulate the inhibitory functions of CTLA-4. These findings have important implications for the use of biologic agents to enhance Treg numbers/functions, as well as for the use of checkpoint inhibitors.
Supported by DIR, NIAID.
Translating knowledge and raising awareness of Immunology related human disease mechanisms via engagement with the general public and lay audience
Authors: Sika-Paotonu Dianne 
Published: January 2023

Abstract
Acute rheumatic fever (ARF) is an autoimmune condition caused by untreated Group A Streptococcal (GAS) infection of the upper respiratory tract (and possibly skin). Multiple or severe attacks of ARF can cause cardiac damage known as rheumatic heart disease (RHD). Effective management for ARF requires painful monthly intramuscular injections of Benzathine Penicillin G (BPG) known as secondary prophylaxis. The goal of secondary prophylaxis is to prevent GAS infections that may lead to the recurrence of ARF. Rates of adherence to secondary prophylaxis are usually low due to the frequency and duration of injection, pain and access to proper and timely healthcare. A less painful and longer acting BPG formulation is urgently needed. Research being undertaken in New Zealand seeks to support global BPG reformulation efforts. The purpose of this work was to support knowledge translation of BPG reformulation research efforts and the associated fundamental Immunology concepts within a public setting. A general public and lay audience outreach event was held through the local university, inviting members of the public to attend. A presentation by a female Pacific Biomedical scientist was offered to support increased knowledge and awareness of emergent rheumatic fever related issues locally and internationally. This event was well attended by the general public. Questions from the audience relating to ARF/RHD dominated, with strong interest expressed in the global efforts to reformulate Penicillin. Additional positive verbal and written feedback was received from those in attendance. This public outreach engagement event supported communication of research findings that was culturally appropriate, clear and meaningful.
Inflammasomes confer protection via IL-18 and pyroptosis, and are negatively regulated by IFN-γ-dependent nitric oxide during Brucella infection
Authors: Lacey Carolyn , Mitchell William J. , Skyberg Jerod A. 
Published: January 2023

Abstract
Brucellosis, caused by the intracellular bacterial pathogen Brucella, is a zoonotic disease for which arthritis is the most common focal complication in humans. Here we show caspase-1 (casp1) and caspase-11 (casp11) initiate Brucella-induced joint inflammation, but later in infection also restrict Brucella replication. In contrast, AIM2 and NLRP3 were dispensable for control of joint infection by Brucella. Both IL-1 and IL-18 promoted joint inflammation, however only IL-18 contributed to protection against Brucella infection of the joint. In vitro studies demonstrated that casp1 and casp11 both induce pyroptosis which limited Brucella infection in macrophages. While IFN-γ production in part required casp1/11 and IL-18, casp1/11-dependent clearance of Brucella was only partially IFN-γ dependent. However, IFN-γ deficiency resulted in severe inflammation that was entirely inflammasome dependent, and in particular reliant on NLRP3. IFN-γ was vital for induction of the nitric oxide (NO) producing enzyme, iNOS, in infected joints, and NO inhibited casp1 activation in Brucella-infected macrophages in vitro. During Brucella infection in vivo, iNOS inhibition exacerbated arthritis, and administration of a NO donor reduced joint inflammation in IFN-γ-deficient mice. Collectively these data demonstrate inflammasomes induce early inflammation in an IL-18 and IL-1 dependent manner, and inflammasome-dependent IL-18 and pyroptosis restrict Brucella burdens in the joint. Moreover, IFN-γ reduces inflammation by inhibiting excessive casp1 activation through production of NO. Thus NO donors, in combination with antibiotics, may be an effective therapy for treating Brucella-induced inflammation.