bluuebunny/crossref_metadata_2025 huggingface dataset:
The α7 nicotinic acetylcholine receptor is an important determinant of monocyte-derived macrophage migration
Authors: Pavlov Valentin A , Cui Kui , Addorisio Meghan , Williams David L , Yakubenko Valentin
Published: January 2023

Inflammation is regulated by the nervous system. The α7 nicotinic acetylcholine receptor (α7nAChR) on macrophages plays a key role in this regulation by meditating cholinergic suppression of pro-inflammatory cytokine release within the vagus nerve-based inflammatory reflex during endotoxemia and other conditions. However, the involvement of of α7nAChR in other macrophage functions remains poorly understood. The objective of this study was to provide insight into the role of α7nAChR in macrophage migration and accumulation in organs during murine endotoxemia. Briefly, monocyte progenitors isolated from bone marrow of wild type (WT) and α7nAChR knockout (KO) mice were labeled with red PKH26 (WT) or green PKH67 (α7nAChR−/−) fluorescent dyes, mixed in equal amounts and injected in tail vein of WT mice 20 min before LPS administration. 48 hours later, lungs, livers and spleens were isolated, digested and analyzed using multicolor flow cytometry (Fortessa-X20) and imaging flow cytometry (ImageStream X Mark II). α7nAChR deficiency resulted in significantly decreased accumulation of macrophages in liver (~10 folds), lung (~4 folds) and spleen (~3 folds) (P<0.01). The morphology of migrated macrophage was verified by imaging flow cytometry. These results identify α7nAChR as an important determinant of the recruitment of monocytes to the site of acute inflammation. They also provide a rationale for further studies on the molecular mechanisms, including the involvement of adhesive receptors in α7nAChR-dependent monocyte/macrophage migration.
In vivo tracking: Using a photoconvertible system to probe immune cell entry and exit in the context of inflammation
Authors: Healey Alicia Marie , Fowell Deborah J
Published: January 2023

Chronic inflammatory diseases such as Crohn’s disease and rheumatoid arthritis affect millions of people worldwide. During an inflammatory response, immune cells enter the inflamed tissue to mediate pathogen clearance, however, their activation and overaccumulation in the tissue can lead to chronic inflammation and autoimmunity. Little is known about the signals that regulate immune cell entry into and exit from these inflamed tissues or about the retention cues that play a role in their accumulation. Therefore, a better understanding of these signals would inform the design of therapeutics intended to alleviate chronic inflammation. To study immune cell movement between a site of inflammation and lymphoid organs, our lab has acquired a mouse model that allows for the in vivo labeling of a subset of cells in time and space. The cells of Kaede transgenic mice constitutively express the photoconvertible fluorescent Kaede protein. CFA/OVA-inflamed ears were photoconverted from Kaede green to red by violet light exposure and the trafficking of Kaede red cells out of the skin to the draining lymph node was assessed over the course of 24 hours. Based on preliminary data, we found that, of the subsets assessed, neutrophils and CD4+ T cells made up the largest percentages of immune cells exiting the inflamed tissue on day 5 of inflammation. On the other hand, far fewer conventional dendritic cells and macrophages were seen to enter the draining lymph node from the inflamed tissue. The role of these tissue-emigrant immune cells is not yet clear, but cells such as neutrophils may play a role in shaping adaptive immune responses in the draining lymph node.
IL-11 induces surface marker expression and migration of T-lymphocytes
Authors: Seyedsadr Maryamsadat , Li Jian , Markovic-Plese Silva
Published: January 2023

Objective

To study the phenotypic and migratory changes of PBMCs following IL-11 stimulation

Background

Multiple sclerosis (MS) is presumably an autoimmune disease in which immune cells mediate neuroinflammation and neurodegeneration in central nervous system. Cytokines play a significant role in orchestrating the inflammatory response during the disease. Our lab has previously shown that in untreated MS patients, IL-11 is upregulated in the CSF and serum in comparison to matched healthy controls (HCs). Previous study has shown that CD4+IL11+ cells are accumulated in the CSF and the brain MS lesions, suggesting that they play a role in the lesion formation.

Methods

14 untreated RRMS patients and age-, sex- and race-matched HCs were enrolled in the study upon signing an Institutional Review Board consent forms.

Results

By using flow cytometry we characterized IL-11-secreting CD4+ lymphocytes and show that they have increased expression of CCR6, CCR4, ICAM-1, IL-11RA, GM-CSF, and IFNγ in comparison to CD4+IL-11− cells (p values<0.03). In vitro studies further showed that stimulation of PBMCs with recombinant human IL-11, upregulated the expression of CCR4 and CCR6 (p values<0.03, 1.2 and 1.4 fold changes) in CD4+ gated cells. In a trans-well system we observed that IL-11 can act as a chemoattractant for anti-CD3/CD28 mAb stimulated PBMCs and increase their migration (p value=0.04, migration index=1.2) in both patients and HCs.

Conclusion

We observed that IL-11 induces migratory phenotype in CD4+ lymphocytes in patients with RRMS, which can be explored as a therapeutic target in early RRMS.
Autologous intraoperative cell salvage ameliorates suppression of transfusion-related monocyte and dendritic cell inflammatory responses
Authors: Dean Melinda M , Obeysekera Maheshi P , Tran Thu V , Sturgess David , Wyssusek Kerstin , Van Zundert Andrea , Gordon Louisa G , Da Silva D , Punnasseril J , Tung John-Paul , Flower Robert L , Roets Michelle
Published: January 2023

ABO compatible allogeneic blood transfusion (ABT) is associated with transfusion-related immune modulation (TRIM) and poor patient outcomes including perioperative infection and myocardial infarction. Intraoperative cell salvage (ICS) enables collection, processing and reinfusion of autologous blood lost during surgery. We investigated whether immune suppression associated with ABT could be reduced by using ICS. Suitable orthopaedic patients were consented and recruited to the study (n=20, HREC/17/QRBW/685). 10mL anticoagulated blood (EDTA) was collected from the patient (pre-operatively) and from the ICS blood product. An established whole blood culture model was used to assess changes to monocyte and myeloid dendritic cell (mDC) inflammatory responses (intracellular cytokine staining for IL-6, IL-8, IL-10, IL-12, IL-1α, TNF-α, MIP-1α, MIP-1β, MCP-1, IP-10) following exposure of the patient’s pre-operative blood to ABO compatible allogenic blood (Blood Service, Kelvin Grove) or their ICS blood (P<0.05 ANOVA). Exposure to ABT significantly suppressed the inflammatory response of both mDC (IL-10, IL-12, IL-1α, IL-8, TNF-a, MIP-α, MIP-1β) and monocytes (IL-10, IL-6, IL-12, IL-1α, IL-8, IL-10, IL-12, TNF-α and MIP-1α). Compared to ABT, exposure to the patients’ own (ICS) blood improved mDC IL-8, IL-10, IL-12, TNF-α and MIP-1α production and monocyte IL-10 and MIP-1β production. We provide in vitro evidence that ICS may reduce ABT associated adverse outcomes associated with improved inflammatory response and immune competence post-transfusion.
Constitutive TGFβ signaling prevents inflammation-induced Langerhans cell migration
Authors: De La Cruz Diaz Jacinto S , Kaplan Daniel H
Published: January 2023

Keratinocytes (KCs) that form the epidermis create a unique barrier niche for cells of the immune system such as Langerhans cells (LCs) and CD8+ resident memory T cells (TRM). These leukocytes provide host-defense but also initiate and maintain autoimmune skin diseases. Hence, elucidating the mechanisms by which CD8+ TRM and LC maintain epidermal residency is of great therapeutic interest. Both cell types require active TGFβ1 to maintain their steady-state epidermal retention. In the epidermis, inactive LAP-TGFβ1 is exclusively activated into active TGFβ1 by integrins ανβ6 and ανβ8 expressed by KCs. However, the relationship between TGFβ1 and LC migration in response to inflammatory stimuli remains unknown. To determine whether enforced TGFβ1 signaling can overcome inflammation-induce LC migration, we generated huLangerin-CreERT2x TGFβRI-CALxL mice (TGFβR1-CALC) that allow for tamoxifen-induced ligand-independent TGFβRI signaling. Enforced TGFβRI signaling in LC prevented migration in response to UVB, chemical haptens, C. albicans epicutaneous infection as well as dermal injection of TNFα and IL1β. LC from TGFβR1-CALC mice did not egress from the epidermis or enter the draining lymph nodes. Notably, in WT mice, we did not observe reduced KC expression of integrin ανβ6 or ανβ8 and levels of pSMAD2 in LC did not decrease in inflammatory contexts. Thus, although enforced TGFβRI signaling can overcome inflammation-induced migratory signals, but the loss of TGFβRI signaling is not required for LC migration. These data are consistent with a model in which TGFβRI signaling drives a LC-intrinsic retention program under homeostatic conditions that is overcome or inhibited by inflammatory cues.
Bactericidal activity of transendothelial neutrophils based the endothelial IL-1α crosstalk during neutrophil–endothelial interaction
Authors: Li Qiuyue , Liu Xiaoye , He Shangwen , Xu Yuanyuan , Zhang Hui , Hou Silu , Dong Hong
Published: January 2023

Neutrophils across endothelial cells to capture and eliminate bacterial pathogens is served as a vocation to prevent infectious diseases during innate immune. However, the mechanisms behind neutrophil-endothelial interaction on the bacterial elimination remains unclear. In this study, we employed a transwell system to co-culture rat intestinal mucosa microvascular endothelial cells (RIMVECs) and neutrophils to illustrate the killing mechanisms of transendothelial neutrophils. We discovered that endothelial interleukin-1α (IL-1α) promoted the survival of rats under Escherichia coli infection. IL-1α enhanced the bactericidal activity of transendothelial neutrophils, while the release of IL-1α is inhibited by lipopolysaccharide (LPS) derived from E. coli during infectious process. In addition, LPS majority damaged RIMVECs on cell membrane and induced cell necrosis. LPS promoted E. coli to escape from neutrophils in vivo and in vitro. Though the isobaric tags for relative and absolute quantification (iTRAQ) method to identify differentially expressed proteins of RIMVECs, we found that IL-1α modulates RIMVECs occurred on the plasma membrane, endoplasmic reticulum and mitochondrial envelope. There were eleven common proteins to persist regulatory. 2 h treatment of IL-1α triggered the cell adhesion molecules (CAMs) of RIMVECs, while oxidative phosphorylation was required for long times’ incubation of IL-1α (4 and 8 h). Our work highlights the critical role of endothelial IL-1α during neutrophil- endothelial interaction, which offered new targets in the bactericidal activity of neutrophils.
Recruitment of kindlin-3 to plasma membrane through its PH domain precedes high affinity β2 integrin activation and neutrophil arrest
Authors: Wen Lai , Marki Alex , Roy Payel , McArdle Sara , Sun Hao , Fan Zhichao , Gingras Alexandre R. , Ginsberg Mark H , Ley Klaus
Published: January 2023

Leukocytes including neutrophils must adhere to the vascular endothelium in order to respond to injury or infection. Integrin-mediated neutrophil adhesion starts by arrest from rolling. Activation of integrins involves conformational changes from an inactive bent conformation to an extended conformation (E+) with high affinity for ligand binding (H+). The cytoplasmic protein kindlin-3 is required for leukocyte adhesion; mutations of kindlin-3 cause leukocyte adhesion deficiency type III syndrome. However, the molecular mechanism by which kindlin-3 regulates β2 integrin activation is unknown. Here we measured the spatiotemporal dynamics of kindlin-3 and β2 integrin conformation changes during neutrophil and HL-60 cell rolling and arrest under flow. Using high-resolution quantitative dynamic footprinting (qDF) microscopy and kindlin-3-fluorescent protein (FP) fusion proteins, we find that kindlin-3 is recruited to the plasma membrane prior to induction of the H+ β2 integrin conformation. Deletion of kindlin-3 or its lipid-binding pleckstrin homology (PH) domain completely abolished H+ β2 integrin induction. Intravital imaging of bone marrow chimeric mice revealed that EGFP-tagged kindlin-3-expressing neutrophils, but not control kindlin-3-deficient neutrophils, arrested in cremaster venules in vivo in response to CXCL1. Our data show that kindlin-3 is indispensable for the H+ β2 integrin conformation and crucial for integrin-mediated leukocyte adhesion during inflammation.
Limited application of IL-2/CD25 leads to persistent IL-2R signaling that promotes gene expression for Treg expansion
Authors: Moro Alejandro , Ban Yuguang , Zhen Gao , Wang Lily , Yu Aixin , Chen Xi , Malek Thomas R
Published: January 2023

Low-dose IL-2 expands Tregs to suppress autoimmunity. However, IL-2 has a short half-life that necessitates frequent administration to maintain increased Treg levels. To improve low-dose IL-2 therapy, we developed an IL-2-based fusion protein, mouse IL-2/CD25, that substantially increases IL-2 half-life to more effectively boost Tregs that controls diabetes in NOD mice. To provide a molecular underpinning for the Treg response to IL-2/CD25, transcriptional profiling was performed by RNAseq for purified Tregs from C57BL/6 mice that received an equivalent amount of IL-2 or IL-2/CD25. Initially, at 4 hr post-injection, differential gene expression analysis revealed a large overlap in significant genes modulated by IL-2 or IL-2/CD25. In striking contrast, at 16 and 72 hr post-injection, only IL-2/CD25 persistently modulated gene expression, with the largest number of IL-2-dependent genes detected at 72 hrs. Canonical pathway analysis revealed many interrelated pathways that control cell growth and signaling. This gene expression pattern by IL-2/CD25, but not IL-2, was also associated with extended STAT5 activation and expansion of Tregs with molecular and phenotypic changes corresponding to activated Tregs. IL-2/CD25 also enhanced gene expression associated with glycolysis and the mitochondrial electron transport chain. Seahorse analysis confirmed that IL-2/CD25 shifted Tregs to a more energetic metabolic phenotype due to an increase in mitocondrial ATP. Collectively, these findings demonstrate that a single injection of IL-2/CD25 persistently stimulates Tregs to activate gene expression to enhance Treg metabolism to fulfill the bio-energetic and biosynthetic demands of activation and proliferation.
Interleukin 7 regulates naive T cell metabolism to promote motility of T cells
Authors: Cannon Judy L , Byrum Janie Rae , Gridley Rebekah , Mrass Paulus , Valenzuela Miriam , Torres David J , Fricke Matthew , Moses Melanie E
Published: January 2023

Interleukin-7 (IL-7) is a key cytokine that drives the survival and maintenance of naïve T cells in lymph nodes. Naïve T cells continuously move throughout lymph nodes in order to interact with dendritic cells (DCs) to become activated and promote an effective immune response. As IL-7 is highly expressed in lymph nodes, we investigated whether IL-7 might regulate T cell movement potentiating T-DC interaction. Using two photon microscopy, we show IL-7 promotes T cell motility in vitro and in vivo. Downstream of IL-7, JAK3 and STAT5 signaling are important to regulate T cell speed. To understand the mechanism underlying IL-7 effects on T cell movement, we investigated whether IL-7 might change the metabolic profile of naïve T cells. Using the Seahorse analyzer, we show that IL-7 can affect the metabolic profile of naïve T cells, particularly CD8 T cells. The chemokine CCL21 has previously been shown to drive T cell motility. We then asked whether IL-7 and CCL21 might combine to modulate effects on T cell metabolism. We find that CCL21 decreases ATP production and this decrease in ATP production persists even in the presence of IL-7. Our results suggest that IL-7 is a novel regulator of T cell motion in lymph nodes via changes in T cell metabolism, leading efficient T cell responses.
Rapid protocol for in vitro induction of cytokine associated with Th2 and Th17 cells from PBMC
Authors: Soto Hortensia , Elia Jeanne , Yu Cuiling , Yu Daniel , Yu Jie , Ni Jessie
Published: January 2023

T helper cell is a key component of the immune system. They mediate humoral and cellular immune responses against pathogens and other diseases like cancer by inducing and activating effector cells such as B cells, cytotoxic T cells and innate cells. They are also involved in the regulation of immune responses by controlling the persistence and magnitude of the response. Activation of naïve T cell is initiated by interaction with antigen presenting cells. The downstream signal transduction and the cytokine milieu can lead to their proliferation and differentiation into different T cell subtypes. To facilitate the study of Th2 and Th17 cells, we present a rapid protocol to efficiently induce cytokine associated with Th2 and Th17 cells from total PBMC. Canonical anti-CD3 and anti-CD28 clones are used to activate the purified cells. Different sets of bioassay-validated cytokines, including IL-12, IL-4, IL-6, IL-1β, IL-2, and TGF-β1 are used to induce cytokine associated with Th2 and Th17 cells. Hallmark cytokine and transcription factor expression profile of different lineages are evaluated by Flow Cytometry. In summary, we have successfully developed a rapid protocols to induce cytokine expression in vitro associated with Th2 and Th17 using high quality reagents developed in BioLegend.
Undergraduate immunology lab project including opportunities for experimental design, a poster presentation, and the opportunity to partially purify antibodies from hybridoma cells to utilize in a Western blot
Authors: Koenigstein Carrie Temple , Johnson Mills
Published: January 2023

Undergraduate students benefit from lab activities that more closely mirror research conditions. In this 7 week lab project, students maintain the hybridoma cell line MARC 2B7 (ATCC CRL-2509), and design a mini-research project to examine antibody production under a modified growth condition of their choice. Lab groups are responsible for making their own solutions, designing controls, and monitoring and maintaining the cell line over a two week period of time. Antibody levels are measured using a dot blot. For each phase, students are presented with a standard lab protocol accompanied by a list of suggestions and/or modifications so that the students are responsible for writing their own final protocol. For example, the dot blot protocol provided is based on chemiluminescent detection; however, the commercial secondary antibody is HRP conjugated so students are also provided the manufacturer’s information on the secondary antibody and the HRP substrate available in lab. This mirrors more real-world lab experiences, and requires students to think more deeply about the experimental design. Each lab group presents a poster on their mini-project to the class. Subsequently, lab groups use ammonium sulfate precipitation followed by a desalting column to concentrate and partially purify the antibodies from hybridoma supernatants that demonstrated positive results on the dot blots. This product is used as the primary antibody for a Western blot at the end of the experiment arc. In addition to providing a platform to discuss both monoclonal and polyclonal antibodies, and enabling students to practice experimental design, this project requires students to utilize their work from early in the semester in a productive final lab experience.
Learning outcomes of using animated video podcasts as a tool for teaching immunology physiology and pharmacology courses in pharmacy curriculum
Authors: Faridi Hafeez , Garcia Ruslan , Castillo Yaimi , Shahid Mohd , Garg Rahul
Published: January 2023

Pharmacy curriculum demands mastery of multiple disciplines such as physiology, immunology and microbiology, medicinal chemistry and pharmacology. Professional pharmacy programs in the United States and other countries have adopted the integrated curriculum model to fulfill the learning objectives. Intensive learning and constant revision of study material is necessary for the success in the pharmacy program. With modern digital technologies and high-speed internet, last decade has witnessed a paradigm shift in traditional pedagogy in every discipline we teach and study. Popularity of online education has grown exponentially in recent years. Several reputed Universities globally now offer blended as well as complete online courses. Large student population from every disciplines now embark on largely available digital resources such as educational videos. As a part of preliminary studies we have prepared few videos and identified some of the potential benefits of using video podcasts in pharmacy curriculum as easy accessibility of material, short video podcast helps better engage students and avoid boredom, animations enhance learning process and memorization, students can study at the ease of watching these videos on phone. Our results suggests strong likability of educational videos as a learning tool among students in pharmacy curriculum.
Teaching immunology laboratory techniques in a course without a lab component using Journal of Visualized Experiments Immunology and Infection, a peer-reviewed scientific journal published in video format
Authors: Walline Crystal C
Published: January 2023

Teaching an undergraduate immunology course without a lab component has many challenges. Firstly, a complicated subject must be taught in about half the number of contact hours. Chiefly, the opportunity to reinforce concepts learned from the instructional setting (e.g. classroom, online module, or in situ teaching) by performing hands-on experimentation is absent. An assignment was devised which required students to read an article published in the Journal of Visualized Experiments (JoVE) Immunology and Infection and watch the companion video. JoVE publishes peer-reviewed detailed experimental techniques in a visual format. Guidelines for critically analyzing a scientific text were provided and students were asked to do a close reading and post questions to an LMS discussion board, which the instructor used to implement the Just-in-Time Teaching (JiTT) method. The assignment was discussed in class by beginning with a list of unfamiliar acronyms and jargon. This provided a starting point for an interactive discussion of the key immunology concepts (e.g. chemotaxis) which was an excellent alternative to a standard lecture. Next, the class watched the video which was paused periodically to address questions posted to the discussion board. An end of discussion assignment was given for students to compare/contrast protein analysis from traditional SDS-PAGE/western blotting to a single-cell, chip-based western blot. Students critically read the literature and learned standard and cutting-edge experimentation/instrumentation in a course without a separate laboratory component. Overall, the use of a visual journal article from JoVE and JiTT led to an interactive, engaging discussion that received positive feedback from students.
Human subjects research as part of immunology education
Authors: Redmond Sarah , Mohamed Attia
Published: January 2023

Original research has been identified as a high impact practice in undergraduate education – students who participate have been shown to be more likely to be be successful in professional and academic graduate programs. Course-embedded research allows for students to tie laboratory experiences to classroom concepts and allows for many students to benefit from faculty oversight. Undergraduate research in immunology presents challenges because most models of immunology are limited to vertebrate organisms, which require additional regulatory oversight from Institutional Animal Care and Use Committees or Institutional Review Boards. One common solution is to limit student interaction with organisms by using isolated or cultured cells, however this prevents students from gaining a realistic understanding of all the stages of immunological research. Obtaining authorization for student-designed research, conducting the projects, and analyzing and presenting the results within a single semester is a daunting proposal, one which has been further complicated by the full implementation of the new Common Rule for human subjects research. We have developed a model for conducting human subjects research within a single semester of a junior-level undergraduate immunology course. By using a framework protocol, non-invasive sampling, and anonymized presented data we are able to work within IRB/IACUC regulations beyond the classroom exercise level, allowing students to share their findings with the broader campus community, in online portfolios, and at professional conferences. This model has been sustainable for five years at a public primarily undergraduate institution and has been successful under the new common rule regulations.
Contextualisation of Immunology related STEM examples to support Youth interest and engagement
Authors: Sika-Paotonu Dianne
Published: January 2023

Efforts are being made to increase the Pacific health workforce in New Zealand (NZ), and also to support the health workforce composition become more reflective of the NZ population. For those embarking upon health based career pathways, this will require participation in STEM and health related subject matter. More work is needed to support the uptake, interest and engagement by Youth in STEM related content – particularly for Pacific students. The purpose of this work was to use STEM related teaching examples with appropriate contextualisation of Immunology based content to support Youth interest and engagement in science amongst Pacific Youth. In-class teaching examples using Cancer related research findings with an Immunological basis were shared with Pacific Youth within group settings that were culturally appropriate and supported group discussion amongst the Youth. These group interactions were facilitated by a Pacific Biomedical Scientist. Positive in-class interactions were supported by the culturally appropriate setting which contributed in supporting favorable student learning and engagement during the classes. Discussion efforts relating to the Cancer related Immunological aspects within these sessions were welcomed by the students, and well-received overall. This work demonstrated the role and importance of cultural considerations needed for Pacific students in supporting positive learning and engagement experiences relating to STEM associated teaching content in the classroom.
Medical immunology for Gen Z
Authors: Geis Abby L , Brown Teressa
Published: January 2023

While there is much discussion in education literature about the benefits of a flipped classroom, medical educators still typically rely on traditional didactic lectures as a primary teaching modality. In fact, when given a choice of delivery modalities, learners often prefer to engage in the easier and comfortable route to knowledge, including listening to lectures. However, traditional learning through passive techniques such as reading and listening is less effective because it fails to facilitate application and recall of concepts. High order learning and retention is significantly improved when the learning process is challenging and thought-provoking.

We use immunology within a medical school curriculum to compare the effectiveness of and student preferences for two distinct teaching modalities. Data will be collected from 170 first year medical students at our institution. The immunology unit consists of 5 sessions, each session linked to 3–4 exam items. Four sessions consist of prerecorded lectures provided to students prior to class. In class, students complete a short individual quiz with low-level questions, the quiz is reviewed, and the remaining class time is spent doing high-level application problems. The final session is delivered as a didactic, in-class lecture. For that final session, students are provided with the low-level quiz and high-level application problems to do at their own discretion. No aspect of the learning activities is required or graded. Following the unit, students will be anonymously surveyed regarding preferences of content delivery modalities. Effectiveness of delivery modalities will be compared by assessing student performance on linked exam items. The unit concludes in November 2019.
Immunology based STEM outreach education efforts that support Knowledge Translation for Pacific College students
Authors: Sika-Paotonu Dianne
Published: January 2023

Specialists and professionals working within STEM related fields can be important role models for students. In the right context and when effective, STEM role models can inspire and motivate children and young people to consider undertaking STEM related classes at school and college. Indigenous Māori and Pacific students in New Zealand (NZ), remain underrepresented within STEM and health related courses and pathways. The importance of role modelling in this particular context has been recognised. An Immunologically based leaning and teaching workshop effort was designed to support a wider STEM-related knowledge translation and outreach education event organised by an external organization. A female Pacific Biomedical Scientist was invited to appear as a STEM role model, and offer a teaching contribution to support increased interest, engagement and participation within the STEM related learning environment. A significant component of the STEM related teaching content was based upon cancer related Immunological content and was tailored for delivery to Pacific College students (aged 14–16 years). The cancer burden amongst Māori and Pacific Peoples in NZ and in the Pacific Region remains of concern. The feedback received from the College students highlighted this session was very well received and enjoyed by the students and was expressed in written and verbal form. Educators can be important role models for their students and for wider knowledge translation efforts. This work demonstrated the importance of role modelling in the classroom when delivering STEM related teaching content for Pacific College students.
Connecting with STEM and Immunology in the Classroom – the importance of Role Modelling
Authors: Sika-Paotonu Dianne
Published: January 2023

Connecting young people with STEM and Immunology within the classroom setting can be a challenge. Outreach education and knowledge translation efforts can be supported by appropriate Role Modelling and Contextualisation for students and young people. The purpose of this work was to support improved interest and engagement by young Pacific students within a STEM related context, within the classroom setting using an Immunology based in-class teaching focus. Pacific students remain underrepresented in STEM and health related subjects and courses. An outreach education STEM based knowledge translational programme was specifically designed for Pacific students aged 10–12 years (Year 7 & 8) by an external organisation, and included an outreach activity contribution by a female Pacific Biomedical Scientist. Context specific STEM related Immunology content was shared with the Pacific students in the form of cancer related research work and was targeted to the 10–12 year age group. The growing burden of cancer remains a significant concern for Pacific Peoples living in New Zealand and the Pacific Region. Student feedback in verbal and written form indicated this in-class session was very well received. Students engaged well with the in-class question and answer session involving the Pacific Biomedical Scientist and expressed interest in considering STEM as possible subject options. The importance of appropriate Role Modelling and Contextualization of STEM associated Immunology based teaching in the Classroom in supporting positive and constructive classroom experiences for students, was reinforced by the feedback and interactions outlined in this work.
Studies of the Properties of a Bouillon Filtrate of the Gonococcus
Authors: Clark L. T. , Ferry N. S. , Steele A. H.
Published: January 2023

It is generally conceded that the gonococcus is a very refractory organism to cultivate artificially, especially in liquid media. When growth does occur in the latter, even in a medium well fortified with any one of the serous human body fluids, the results are extremely uncertain, particularly in amount and duration of proliferation. This lack of adaptability to artificial cultural conditions perhaps accounts for the comparatively little work that has been reported in the literature referring to systematic studies on germ-free filtrates of the gonococcus and methods suited to their preparation and evaluation.

The inherent tendency of cultures of the gonococcus to autolyze offers another obstacle to the analysis of such filtrates, as found by McClintock and Clark (1) and others.

The work presented in this paper covers the preparation and testing of a toxic filtrate from liquid culture of the gonococcus which followed a request from Dr. B. C. Corbus of Chicago for a bouillon filtrate of the gonococcus in the nature of an “antivirus.”
On the Forssman Antigens in B. Paratyphosus B and B. Dysenteriae Shiga
Authors: Landsteiner K. , Levine Philip
Published: January 2023

The occurrence of substances in bacilli of the Salmonella group,1 inducing in rabbits the formation of sheep hemolysins, so-called Forssman antibodies, has been described by various authors (4), (5), (6), (7).2 Such antigens have been found by Seiffert (6) in strains of B. paratyphosus (Schottmüller and Freiburg) and of B. enteritıdis Gaertner, and by Meyer in the types Newport and Berlin. Other bacilli which have been shown to contain Forssman antigens, are B. dysenteriae Shiga (9), (10), a particular strain of B. leptisepticus (11), and pneumococci (12), (13).

That the various bacillary antigens are not identical, has been shown by Jungeblut and Ross for B. paratyphosus B and B. dysenteriae Shiga, and Meyer has demonstrated that immune sera containing sheep lysin, produced by four different strains of the Salmonella group (B. paratyphosus B, B Berlin, B Newport, and B. enteridıtis Gaertner) were absorbable only by the homologous organisms (see Eisler (14)).
A Contribution to the Study of the Etiology of Serum Disease
Authors: Meyers H. Russell
Published: January 2023

Serum reactions following the parenteral introduction of cock serum appear to possess characteristics distinct enough from serum disease following injections of horse serum to warrant a differentiation. Horse serum fibrinogen does not appear to be a responsible etiologic agent in the production of serum disease, for: When prepared reasonably free from other protein elements and injected intravenously in an amount equivalent to or greater than the amount estimated to be contained in a quantity of horse serum capable of producing serum disease in 90 per cent of patients, fibrinogen did not produce serum sickness in the series reported here. The concentrated sera prepared according to the Atkinson-Banzhaf method by the Department of Health in the City of New York does not contain detectable fibrinogen, although these sera are known to cause serum disease.
Type I IFN Receptor Signaling on B Cells Promotes Antibody Responses to Polysaccharide Antigens
Authors: Spurrier M Ariel , Jennings-Gee Jamie E , Haas Karen M
Published: January 2023

We previously reported monophosphoryl lipid A (MPL) and synthetic cord factor trehalose-6,6′-dicorynomycolate (TDCM) significantly increase Ab responses to T cell–independent type 2 Ags (TI-2 Ags) in a manner dependent on B cell–intrinsic TLR4 expression, as well as MyD88 and TRIF proteins. Given the capacity of MPL to drive type I IFN production, we aimed to investigate the extent to which type I IFN receptor (IFNAR) signaling was required for TI-2 responses and adjuvant effects. Using Ifnar1−/− mice and IFNAR1 Ab blockade, we found that IFNAR signaling is required for optimal early B cell activation, expansion, and Ab responses to nonadjuvanted TI-2 Ags, including the pneumococcal vaccine. Further study demonstrated that B cell–intrinsic type I IFN signaling on B cells was essential for normal TI-2 Ab responses. In particular, TI-2 Ag–specific B-1b cell activation and expansion were significantly impaired in Ifnar1−/− mice; moreover, IFNAR1 Ab blockade similarly reduced activation, expansion, and differentiation of IFNAR1-sufficient B-1b cells in Ifnar1−/− recipient mice, indicating that B-1b cell–expressed IFNAR supports TI-2 Ab responses. Consistent with these findings, type I IFN significantly increased the survival of TI-2 Ag–activated B-1b cells ex vivo and promoted plasmablast differentiation. Nonetheless, MPL/TDCM adjuvant effects, which were largely carried out through innate B cells (B-1b and splenic CD23− B cells), were independent of type I IFN signaling. In summary, our study highlights an important role for B-1b cell–expressed IFNAR in promoting responses to nonadjuvanted TI-2 Ags, but it nonetheless demonstrates that adjuvants which support innate B cell responses may bypass this requirement.
Low-Dose Lipopolysaccharide Protects from Lethal Paramyxovirus Infection in a Macrophage- and TLR4-Dependent Process
Authors: Resiliac Jenny , Rohlfing Michelle , Santoro Jennifer , Hussain Syed-Rehan A , Grayson Mitchell H
Published: January 2023

Respiratory diseases are a major public health burden and a leading cause of death and disability in the world. Understanding antiviral immune responses is crucial to alleviate morbidity and mortality associated with these respiratory viral infections. Previous data from human and animal studies suggested that pre-existing atopy may provide some protection against severe disease from a respiratory viral infection. However, the mechanism(s) of protection is not understood. Low-dose LPS has been shown to drive an atopic phenotype in mice. In addition, LPS has been shown in vitro to have an antiviral effect. We examined the effect of LPS treatment on mortality to the murine parainfluenza virus Sendai virus. Low-dose LPS treatment 24 h before inoculation with a normally lethal dose of Sendai virus greatly reduced death. This protection was associated with a reduced viral titer and reduced inflammatory cytokine production in the airways. The administration of LPS was associated with a marked increase in lung neutrophils and macrophages. Depletion of neutrophils failed to reverse the protective effect of LPS; however, depletion of macrophages reversed the protective effect of LPS. Further, we demonstrate that the protective effect of LPS depends on type I IFN and TLR4-MyD88 signaling. Together, these studies demonstrate pretreatment with low-dose LPS provides a survival advantage against a severe respiratory viral infection through a macrophage-, TLR4-, and MyD88-dependent pathway.
Inefficient Recovery of Repeatedly Stimulated Memory CD8 T Cells after Polymicrobial Sepsis Induction Leads to Changes in Memory CD8 T Cell Pool Composition
Authors: Moioffer Steven J , Berton Roger R , McGonagill Patrick W , Jensen Isaac J , Griffith Thomas S , Badovinac Vladimir P
Published: January 2023

Long-lasting sepsis-induced immunoparalysis has been principally studied in primary (1°) memory CD8 T cells; however, the impact of sepsis on memory CD8 T cells with a history of repeated cognate Ag encounters is largely unknown but important in understanding the role of sepsis in shaping the pre-existing memory CD8 T cell compartment. Higher-order memory CD8 T cells are crucial in providing immunity against common pathogens that reinfect the host or are generated by repeated vaccination. In this study, we analyzed peripheral blood from septic patients and show that memory CD8 T cells with defined Ag specificity for recurring CMV infection proliferate less than bulk populations of central memory CD8 T cells. Using TCR-transgenic T cells to generate 1° and higher-order (quaternary [4°]) memory T cells within the same host, we demonstrate that the susceptibility and loss of both memory subsets are similar after sepsis induction, and sepsis diminished Ag-dependent and -independent (bystander) functions of these memory subsets equally. Both the 1° and 4° memory T cell populations proliferated in a sepsis-induced lymphopenic environment; however, due to the intrinsic differences in baseline proliferative capacity, expression of receptors (e.g., CD127/CD122), and responsiveness to homeostatic cytokines, 1° memory T cells become overrepresented over time in sepsis survivors. Finally, IL-7/anti–IL-7 mAb complex treatment early after sepsis induction preferentially rescued the proliferation and accumulation of 1° memory T cells, whereas recovery of 4° memory T cells was less pronounced. Thus, inefficient recovery of repeatedly stimulated memory cells after polymicrobial sepsis induction leads to changes in memory T cell pool composition, a notion with important implications in devising strategies to recover the number and function of pre-existing memory CD8 T cells in sepsis survivors.
Precipitation Test for the Sexes of Fowl Blood Serum, with Special Reference to Egg-Laying
Authors: Sasaki Kiyotsuna
Published: January 2023

IV. Summary

Injecting the rabbits with the fowl blood sera, the sex tests of the blood sera were made by means of precipitin reaction, with special reference to egg-laying, and the results are as follows: The blood serum of the laying hen can not be distinguished from that of the cock or of the non-laying hen with the corresponding antiserum. However, by specialization of the antiserum, the former is capable of being clearly distinguished from the two latter with the absorbed antiserum for the blood of the laying hen, a specific reaction for the homologous serum being given; whereas sera of the cock and the non-laying hen are not distinguishable from each other with the corresponding absorbed antisera.The blood serum of the pullet gives similar results with that of the cock or of the non-laying hen. Therefore, the blood sera of immature fowl, namely, pullet, and those of mature ones, excepting the laying hen, can not be distinguished mutually.The antiserum for laying hen's blood serum contains specific precipitin for egg-laying not for sex.
Further Attempts to Transfer the Immunological Response of Tuberculous Animals
Authors: Dienes L.
Published: January 2023

Summary and Conclusions

Three groups of experimental efforts to transfer to other animals the increased immunologic response of tuberculous animals have been described. The transfer of tuberculous omenta or tuberculous peritoneal exudate, a few minutes after the injection of egg-white, to the peritoneal cavity of normal pigs was unsuccessful. Where a similar transfer was made into the peritoneal cavity of a previously infected animal—in a stage of infection in which the injection of egg-white alone does not produce the characteristic effect—irregular positive results were obtained. A simultaneous injection of egg-white and tubercle bacilli was in these guinea pigs often successful.

The experiments suggest that two factors are involved, (a) the effect which the lesion itself produces upon the antigen, and (b) a certain preparation of the organism as a whole occurring during the course of the infection.
Serological Diversity Among Meningococci
Authors: Branham Sara E.
Published: January 2023

Accurate “typing” of meningococci is often a laborious process. This is due chiefly to the following factors: (a) Lack of specificity among meningococci; (b) lack of specificity in “typing” serum; (c) lack of antigenic stability in meningococci; and (d) failure to do the “typing” soon after isolation. It is frequently necessary to use a combination of many methods in order to place some strains in their respective serological groups.

The relative proportion of meningococci which fall into the four groups of Gordon and Murray has not been the same during the 1927–1930 outbreaks as during 1915–1918. More than 80 per cent of strains studied have been placed in the I and III groups, the relation of which to each other is so intimate that the classification of many strains as I or III has been difficult.

Intensive study of the serological pattern of meningococci is important (a) in respect to the choice of strains to use in making therapeutic sera, and (b) in choosing standard “type” strains. The value of expending the amount of time and labor necessary for the satisfactory classification of complex strains in some routine work may be questioned.
The Mouse Protection Test on Antipneumococcic Serum Type I
Authors: Beard Stanley D. , Clapp Frances L.
Published: January 2023

In evaluating the potency of antipneumococcic sera, the mouse protection test, based on the work of Neufeld (1) and that done at the Rockefeller Institute (2), has been extensively used, and has been prescribed by the Public Health Service (3), for the standardization of antipneumococcic serum type I. The minimum standard of potency as established in the official test is that 0.2 cc. of serum must protect at least 50 per cent of the mice injected against 0.1 cc. of virulent type I culture, this dose of culture being equivalent to about 10 million fatal doses of culture.

The New York State minimum standard (4) is essentially the same as the of the Public Health Service, but in actual practice the test has been somewhat modified. For example, 0.1 cc. of serum is often used in the serum-culture mixtures injected in the test mice as well as 0.2 cc.; also, 0.2 and 0.3 cc. doses of culture are used as well as the 0.1 cc. dose.
Studies on the Immunizing Substances in Pneumococci
Authors: Felton Lloyd D.
Published: January 2023

Pneumococci, both type I and type II, dried by application of acetone followed by vacuum desiccation, retain their antigenicity.

The cell may be divided into acid soluble and acid insoluble constituents. The acid soluble fraction possesses most of the immunizing activity of the cell; the immunity obtained by injecting this fraction into white mice is largely specific. The acid insoluble fraction, perhaps containing some intact cells, also possesses a small amount of the immunizing substance and produces in white mice a heterologous immunity.

With the method used in this study, at least 90 per cent of the immunizing substance in the acid soluble fraction may be precippitated by ethyl alcohol or by acetone.

The work here reported is limited to tests on white mice, and to antigenicity as produced by a single injection of the immunizing substance.
Age and Antibody Production
Authors: Baumgartner Leona
Published: January 2023

It has previously been demonstrated by us that the avidity of fresh antisera agglutinating B. enteritidis produced by the active immunization of young rabbits is greater than that of antisera produced in adult or aged animals. Additional observations on these sera led to the following conclusions: These avidity differences diminish with the “aging” in vitro of the antisera.Antisera partially separated from serum proteins by adsorption to kaolin and elution with glycocoll, exhibit the same trends in avidity differences as do fresh sera.The temperature at which the observations were made did not affect the reaction.Antisera agglutinating B. enteritidis obtained from animals of various ages exhibit similar avidity differences when tested against Eb. typhosus, that is, minor agglutinins exhibit the same qualities as do major agglutinins.In antisera produced by the injection of a bivalent antigen composed of sheep erythrocytes and B. enteritidis, the hemolysin produced by the adult animal shows a greater avidity for its antigen than does the hemolysin produced by young or very aged animals.The avidity of a given antiserum is a property characteristic of that serum, and not directly related to its titer.