bluuebunny/crossref_metadata_2025 huggingface dataset:
Development of therapeutics to maintain vaccine immunity following exposure to ionizing radiation.
Authors: McFarland Hugh I , Surujdin Ryan T , Aryankalayil Joseph , Berkson Julia D. , Lee Jay P. , Rosenberg Amy S.
Published: January 2023

Exposure to γ radiation from nuclear power plant disasters, terrorist acts, and medical procedures devastate the peripheral immune system, killing both naïve and memory T cells. Although, naïve T cells recover, memory T cells mediating vaccine immunity are permanently lost rendering a victim highly susceptible to infection. There is no treatment to globally preserve vaccine immunity following ionizing radiation exposure. We have recently published that CD8+ memory T cell responses to Listeria can be preserved in the aftermath of radiation exposure by revaccination within 3–4 days after irradiation. A correlation between survival and T cell activation prior to irradiation has been demonstrated in vitro. Comprehensive revaccination in a radiation mass casualty situation would be problematic from many perspectives. We are working to develop a therapeutic regimen to replicate the pro-survival effects of revaccination. Among the signaling pathways stimulated by T cell activation is the Akt “survival pathway” which inhibits apoptosis, and enhances the rate and fidelity of repair of damaged DNA. Agonists or inhibitors targeting the various receptors and signaling molecules that interact within this pathway may result in survival of memory T cells and are potential therapeutic candidates. In addition, revaccination rescue upregulates p21 mRNA. p21 is strongly anti-apoptotic, and enhances DNA strand break repair. In preliminary studies, a number of candidate therapeutics have been identified including nicotine, which virtually eliminates splenic Listeria in immunized and irradiated mice following challenge with wild-type bacteria. Testing is underway with nicotine patches which would provide an ideal method of administration.
Radiation Therapy and Talimogene Laherparepvec (TVEC) Synergize in the Treatment of Melanoma
Authors: Jhawar Sachin R , Goyal Sharad , Thandoni Aditya , Bommareddy Praveen , Hassan Suemair , Schiff Devora , Haffty Bruce G , Kaufman Howard , Zloza Andrew
Published: January 2023

Talimogene laherparepvec (TVEC), an oncolytic virus, is an FDA-approved therapy that increases the overall survival of locally advanced and metastatic melanoma patients. Radiation therapy (RT) in combination with immunotherapies improves response rates (compared to either modality alone). Therefore, we hypothesized that combination RT and TVEC results in a synergistic therapeutic effect on melanoma. We treated 8 human melanoma cell lines with radiation (0, 4, or 8 Gy) and/or T-VEC (0, 0.01, 0.1, or 1 MOI) and determined its therapeutic effects using the AlamarBlue cell viability assay. Treatment of UACC257 melanoma cells with combination RT (8 Gy) and TVEC (1 MOI) resulted in a significant decrease in tumor cell viability (a decrease of 44.9%) compared to no treatment, RT alone (33.2%, P=0.0004), or T-VEC alone (24.2%, P=0.0088), with similar findings in other cell lines. We then injected a B16 mouse melanoma cell line, permissive to T-VEC infection, into B6 mice to assess in vivo synergy. Mice were treated when tumors were ~7mm × 7mm. RT resulted in an early 20% decrease in tumor size within 4 days, followed by an increase thereafter. Intra-tumoral TVEC resulted in no clear benefit until 9 days post-treatment, when it showed a slight decrease in size sustained for ~10 days. Combination treatment, however, led to an early response that was sustained until 18 days after the initial treatment. When tumors were assessed 18 days after initial treatment, a 3-fold or more increase in tumor size was lowest in the combination T-VEC + RT group (17%) compared to the control (100%), RT (60%), or TVEC (50%) groups. These data propose that combination T-VEC + RT may be beneficial for cancer patients, and studies establishing the mechanism of synergy are ongoing.
RNAdjuvant®, a novel, highly-potent RNA-based adjuvant, combines strong immunostimulatory capacities with a favorable safety profile
Authors: Lutz Johannes , Heidenreich Regina , Mehr Keyvan Tadjalli , Noth Janine , Koch Sven D , Döner Fatma , Hong Henoch S , Melber Karl , Dähling Angelika , Roos Tilmann , Kowalczyk Aleksandra , Baumhof Patrick , Scheel Birgit , Voss Söhnke , Kallen Karl-Josef , Fotin-Mleczek Mariola , Gnad-Vogt Ulrike
Published: January 2023

Purified recombinant proteins and peptides, which are currently under development in various anti-cancer vaccination approaches, lack sufficient immunogenicity. Therefore, potent adjuvants are needed to induce strong and persistent anti-tumor immunity. However, currently only few adjuvants are licensed, most of which primarily enhance antibody, but not T cell responses.

Here, we demonstrate that a novel, well defined, and thoroughly characterized RNA-based adjuvant mediates balanced and long-lasting humoral and cellular immune responses. Our adjuvant significantly enhances anti-tumor immunity, and even complete tumor rejection can be achieved as shown for the syngeneic TC-1 tumor model, a murine model of human HPV-induced cervical cancer.

Our adjuvant acts locally, promoting strong but transient up-regulation of anti-viral and pro-inflammatory cytokines, CXCR3-ligands and cytoplasmic RNA sensors at the injection site, avoiding any systemic cytokine release. A phase I first in man clinical trial testing different doses of RNAdjuvant® alone and in combination with reduced doses of the licensed rabies vaccine Rabipur® was successfully conducted in 43 subjects. Healthy volunteers received 2 intramuscular injections of RNAdjuvant® on days 0 and 21, either alone or in combination with 1/20 or 1/10 of the licensed Rabipur® dose. Virus neutralizing antibody titers (VNTs) measured on days 14 and 28 revealed a significant increase in median VNTs in subjects with RNAdjuvant® compared to their respective control group with 1/10 dose Rabipur® alone.

In summary, our data suggest that RNAdjuvant® represents a novel, highly efficacious adjuvant candidate that can enhance cellular and humoral immune responses.
Revealing the mechanism of regulatory T-cell generation in vivo by novel Fluorescent Timer reporter
Authors: Ono Masahiro , Bending David , Martin Paz Prieto
Published: January 2023

Foxp3+ regulatory T-cells (Treg) are considered to mainly differentiate in the thymus and protect against autoimmunity. However, the physiological significance of peripheral Treg differentiation is still unclear, due to a lack of methods to identify newly generated Treg in vivo. In addition, the mechanism of Treg differentiation in the thymus is still obscure, because of difficulties in defining temporal sequences of developmental events in vivo.

Here we develop a ground-breaking technology to reveal the time-dependent mechanisms of Treg generation and maturation in vivo. Using Fluorescent Timer protein that spontaneously and irreversibly changes its emission spectrum, we establish a novel Foxp3Timer reporter, and and visualize the dynamics of Foxp3 transcription and thereby identify de novo Foxp3 expression in vivo. Here we show that antigenic recognition immediately induces de novo Foxp3 expression in activated effector T-cells in vivo. Surprisingly, most of Foxp3-expressors in inflamed tissue are not derived from the pre-existing Treg pool, but are newly generated upon antigen recognition. RNA-seq analysis reveals novel genomic mechanisms for peripheral Treg generation and identifies new markers for antigen-reactive Treg, by which we successfully manipulate Treg differentiation dynamics.

Collectively, we show that Foxp3 constitutes a negative feedback mechanism for activated effector T-cells, which induces the resolution of inflammation. Our novel Timer technology is applicable to many outstanding problems in immunology.
Controlling T cell development and function via innate Toll-like receptor signaling for autoimmune therapy
Authors: Liu Xia , Ye Jian , Wang Yadan , Zhong Xiaoli , Li Lingyun , Opejin Adeleye , Hsueh Eddy C. , Luo Huanle , Wang Tian , Hawiger Daniel , Peng Guangyong
Published: January 2023

Innate regulation through Toll-like receptor (TLR) signaling has been shown to be important for promoting T cell subset development and function. However, limited information is known about whether differential TLR signaling can selectively inhibit Th17 and/or Th1 cells, important for controlling excessive inflammation and autoimmune responses. Here we demonstrate that activation of TLR7 signaling in T cells can inhibit both Th17 cell differentiation from naïve T cells and IL-17 production in established Th17 cells. We further report that down-regulation of STAT3 signaling is responsible for TLR7-mediated inhibition of Th17 cells due to induction of SOCS3 and SOCS5. TLR7-mediated suppression of Th17 cells does not require DC involvement. In addition, we show that TLR7 signaling can suppress Th1 cell development and function due to a mechanism different from Th17 cell suppression. Importantly, our complementary in vivo studies demonstrate that treatment with the TLR7 ligand imiquimod can inhibit both Th1 and Th17 cells, resulting in prevention of and immunotherapeutic reduction in experimental autoimmune encephalomyelitis (EAE). These studies identify a new strategy to manipulate Th17/Th1 cells through TLR7 signaling, with important implications for successful immunotherapy against autoimmune and inflammatory diseases.
Unique interaction dynamics and peptide-MHC class II (pMHC II) transendocytosis lead to antigen-specific T regulatory cell (Treg)-mediated suppression
Authors: Akkaya Billur , Akkaya Munir , Kamenyeva Olena , Kabat Juraj , Dorward David W , Holstein Amanda H , Maz Mitra P , Shevach Ethan M
Published: January 2023

Tregs can employ numerous mechanisms for inhibiting immune responses. Antigen-specific Tregs exhibit enhanced capacity to suppress compared to polyclonal Tregs, but unique suppressor mechanisms used by antigen-specific Tregs have not been elucidated. We first compared the interaction of antigen-specific induced Tregs (iTregs) and antigen-specific T effector cells with dendritic cells (DC) using scanning electron microscopy and intravital two-photon microscopy. Antigen-specific iTregs uniquely and rapidly formed dense clusters around DC suggesting that they may prevent the access of antigen-specific CD4+ T cells to the DC surface. In an adoptive transfer model, this rapid interaction was accompanied by a decreased ability of co-transferred naïve T cells to interact with the DCs as indicated by their failure to decrease their motility. More importantly, antigen-specific iTregs were found to specifically transendocytose cognate peptide-MHCII complexes from the DC surface to a greater extent than T effector cells resulting in diminished levels of antigen on the DC surface. Transmission electron microscopy of DC-Treg co-cultures showed that Tregs engulfed parts of DC processes as membrane invaginations within two hours and then internalized the p-MHCII complexes into endosomal vesicles. Taken together, these results indicate a two-step process by which antigen-specific Tregs inhibit antigen presentation. They rapidly and efficiently adhere to the DC surface and then deplete the pMHC II complexes from the DC resulting in potent suppression of the capacity of the DC to activate antigen-specific T cells.
Mer tyrosine kinase (MerTK) deficiency drives the loss of marginal zone macrophages with subsequent accumulation of apoptotic cells and dysregulation of germinal centers
Authors: Soni Chetna , Schell Stephanie L. , Fasnacht Melinda , Rahman Ziaur S.M.
Published: January 2023

MerTK is a scavenger receptor involved in anti-inflammatory efferocytosis. We previously reported that MerTK deficiency on a mixed B6 and 129 background (B6-129.MerKO) led to dysregulated germinal center (GC) responses and accumulation of apoptotic cells (ACs), primarily in the GCs. Here we report that the dysregulation of GCs in B6-129.MerKO mice strongly correlates with the loss of marginal zone macrophages (MZMs) starting at two months of age. In addition to MerTK, MZMs also express other AC-scavenger receptors like SIGNR1, MARCO and Tim-4, which are also lost from the splenic MZ of B6-129.MerKO mice. We do not observe a loss or translocation of marginal zone B cells (MZBs) or marginal zone metallophillic macrophages (MMMs) from the marginal zone, indicating the exclusive role of MZMs in deregulated GC responses observed in this model. Preliminary results using splenic and bone marrow derived macrophages show reduced expression of a pro-survival heat shock protein Hsp27 and its phosphorylation through p38 and MAPKAPK-2 in the absence of MerTK. We also observe increased cell death in MerTK deficient macrophages vs. WT macrophages, when challenged with ACs. Our data indicate that reduced Hsp27 expression due to MerTK deficiency may lead to cell death of MZMs.

We are further investigating if the increased availability of self-antigens through AC accumulation, due to the loss of MZMs can lead to positive selection of self-reactive B cells in the GCs, causing autoimmunity. Our data indicate that MerTK deficiency and ACs enhance antigen presentation by macrophages, DCs and B cells. In accordance, we observe an increase in activated T cells in MerTK deficient mice, which may promote dysregulation of GC responses observed in this model.
Bright blue fluorescent nanoparticles for live cell imaging by fluorescence microscopy
Authors: Richey Kristen H , Zhuang Meng , Fraser Cassandra L. , Pompano Rebecca R.
Published: January 2023

Fluorescence imaging of labeled cells is an important method to monitor long-term cell behaviors. Long-lasting green stains such as CFSE (carboxyfluorescein succinimidyl ester) and its derivatives are routinely utilized for cell migration studies, but currently there are no commercially available long-lasting (days), non-cytotoxic blue cell stains compatible with a standard DAPI filter that is brighter than the autofluorescence of tissue. Here, we fabricated two bright blue emitting boron dye-polymer nanoparticles (BNPs) (e = 32100 M−1cm −1) from difluoroboron dibenzoylmethane conjugated to poly(lactic acid) (BF2dbmPLA) or poly(e-caprolactone) (BF2dbmPCL). Particles were < 100 nm diameter, and their excitation and emission wavelengths (lex = 381 nm; lem = 439 nm) corresponded to the DAPI filter. When incubated with mouse splenocytes, the blue emitting nanoparticles were taken up by the cells and readily detected by fluorescence microscopy (EVOS microscope) or a plate reader. Viability of the stained cells was assessed by calcein/propidium iodide staining. These data showed that BNPs have the capability to stain cells brightly without significantly diminishing viability and therefore show promise as a new tool for live cell tracking in the blue fluorescence channel. We are currently measuring cell fluorescence and viability over time, determining which types of cells take up the particles, and comparing the staining capabilities of the nanoparticles to commercial blue dyes.
CD47 antibody-induced engulfment of human T cell leukemia cells by bone marrowderived macrophages
Authors: APPLEDORN DANIEL , Lovell Gillian , Szybut Clare , Patel Kalpana , Campwala Hinnah , Bevan Nicola , Dale Tim , Trezise Derek
Published: January 2023

CD47 is a trans-membrane “don’t-eat-me” signaling protein that enables tumor cells to evade clearance by neighboring phagocytes. Blocking CD47 allows phagocytes to clear tumor cells and is a promising new approach for cancer immunotherapy. In this study, we characterized anti-CD47 antibody-mediated engulfment of living tumor cells (CCRF-CEM) by mouse bone-marrow derived macrophages (BMDMs) or immortalized mouse macrophages (J774A.1). Phagocytosis was quantified using a pH-sensitive cell-labeling fluorescent probe, pHrodo, and automated kinetic live-cell analysis (IncuCyte). CCRF-CEM cells were labeled, washed and then treated with antibody. Target cells were then added to BMDMs or J774A.1 cells in 96-well plates. Phase- and fluorescence images were captured and quantified over time. Anti-CD47 antibody, but not IgG-control, produced time- and concentration-dependent engulfment of CCRF-CEMs by BMDMs (30’-4h), as evidenced by a significant increase in intracellular fluorescence. Close inspection of the time-lapse images revealed clear cellular engulfment coincident with the appearance of the fluorescent signal. Similar observations were made with J774A.1 macrophages. Interestingly, the rate and degree of engulfment was effector cell-dependent. The mechanism of engulfment was not via induction of target cell apoptosis since anti-CD47 did not induce PS externalization (Annexin V) or activate caspase 3/7. Our experimental findings substantiate the known pro-phagocytic effects of anti-CD47 antibodies, and provide a model system and method for quantitative functional analysis and mechanistic insight of CD47 modulators as cancer therapeutics.
Analysis of Mixed Microbial Populations and Deep Immunotyping of Peripheral Blood on the ZE5 Cell Analyzer
Authors: Green Angela M , Scott Michelle , Kroeger Kelly
Published: January 2023

With up to 5 lasers and 30 analysis parameters, the ZE5 flow cytometer is equipped for complex multiplex analysis of cells. Excellent fluorescence sensitivity (<100MESF values in FITC, PE and APC channels) enables detection of antigens expressed at low levels. The three scatter options; forward scatter (FSC), side scatter (SSC) and forward scatter small particle detector (FSCspd) provide optimal resolution of particles >0.3um to 50um in size. Furthermore, the option to set a dual trigger using any scatter or fluorescence channels allows for optimal background reduction. For this study, the ZE5 was used to evaluate size beads and mixed microbial populations, and to perform immunophenotyping of multiple cellular subsets in human blood using multi-color panels. Beads sized 0.3um and larger were discriminated from noise when using scatter or fluorescence as a trigger. Analysis of mixed microbial populations clearly identified the unique populations within the mixture. For immunophenotyping experiments, PBMCs from normal healthy donors were stained using a 14 color panel to identify T cell, B cell, NK cells, dendritic cells and monocyte sub populations. The panel data was compensated accurately on the ZE5 and each sub-cellular population was well identified.
Development and testing of a novel multiplex serodiagnostic assay for Zika and other arboviruses
Authors: Venkateswaran Neeraja , Sarwar Jawad , Parameswaran Nishnath , Fecteau Tracy , O’Connor David , Nelson William M , Venkateswaran Kodumudi
Published: January 2023

Human immunoglobulin (IgM) antibody capture enzyme linked immunosorbent assay (MAC-ELISA) is recommended by US Centers for Disease Control (CDC) to detect the Zika Virus (ZIKV) infection in samples collected after few days of the onset of disease. However, the cross-reactivity of IgM and immunoglobulin G (IgG) antibodies to ZIKV with dengue virus (DENV), West Nile virus (WNV), yellow fever virus (YFV) and some other related flaviviruses poses a significant challenge for an accurate detection of ZIKV infection in a singleplex serological assay. To overcome these challenges we developed a novel multiplex serological assay that will simultaneously detect antibody response to ZIKV, other related flaviviruses and Chikungunya virus (CHIKV). Various recombinant arboviral antigens were coupled to optically-coded microspheres from Luminex Corporation. This fluorescent labeled microsphere assay was used to differentiate between recent, past arbovirus infections, or co-infections that may occur in endemic regions. We used plasma samples from non-human primates (NHPs) prior to and after ZIKV infection for longitudinal assessment of IgM and IgG antibody responses for evaluation of this method. 37 blood samples from 5 different animals challenged with either African strain or Asian lineage strain of ZIKV were analyzed for antibody responses to multiple antigens included in this test. We developed a detection algorithm for automation of IgM result analysis to differentiate ZIKV IgM response from other arboviruses. Thus this multiplex arbovirus assay has great promise for testing human samples for detecting ZIKV infection.
Control of alloreactivity using transient expression of a monoclonal antibody directed chimeric antigen receptor in conventional and regulatory T cells
Authors: Pierini Antonio , Iliopoulou Bettina , Peiris Heshan , Perez-Cruz Magdiel , Baker Jeanette , Hsu Katie , Strober William , Zheng Ping Ping , Erkers Tom , Alvarez Maite , Ring Aaron , Velardi Andrea , Negrin Robert S. , Kim Seung , Meyer Everett
Published: January 2023

The genetic modification of T cells offers many promising cellular therapies, but little known about how the transient targeting of T cell or T cell subsets such as CD4+CD25+FoxP3+ regulatory cells (Treg) could be used to control alloreactivity. We used the expression of a chimeric antigen receptor that recognizes labeled therapeutic monoclonal antibodies, to control T cell activation in vivo. By specifying the location of activation of donor T cells during hematopoietic cell, we modulated graft-versus-host disease and anti-leukemia effects. We also controlled the activation of Treg cells which remain immunoregulatory. When recipient Treg are targeted to activate in response to MHC Class I mismatch, we observed significantly prolonged allogeneic islet graft survival in an antigen-specific manner, as secondary skin grafts were specifically protected against rejection. The transient genetic modification of T and Treg can have lasting effects on the immune system and could be used therapeutically to regulate alloreactivity.
Bone marrow-derived mesenchymal stem cells (MSCs) to enhance nerve regeneration and functional recovery in vascular composite allotransplantation (VCA)
Authors: Salgar Shashikumar K , Bingham Jason , Kniery Kevin , Jorstad Nikolas , Horkayne-Szakaly Iren , Hoffer Zachary
Published: January 2023

Introduction

Optimizing nerve regeneration and mitigating denervation atrophy of target muscles are the keys to achieving successful VCA such as limb and face transplants.

Methods

Lewis rat was anesthetized, sciatic or individual sciatic nerve branches (tibial, peroneal and sural) of the right hind-limb were transected and surgically repaired. MSC (5 million; passage ≤6) or vehicle were administered locally and intravenously.

Results

Rat MSCs expanded ex vivo were CD29+, CD90+, CD34−, CD31−, CD45low, MHC Class I+, Class II−, and were pluripotent. Total sensory function recovery at 2 weeks post sciatic or individual nerve repair (SNR or INR) with or without MSC was ~ 1.2 on a scale of Grade 0–3 (0=No function; 3=Normal); by 12 weeks it was 2.6–2.8 (n≥6/group). Peroneal sensory function recovery was as early as 1 week but not tibial or sural. MSC treatment accelerated sensory function onset. At 8 weeks post-INR, the sciatic nerve function index (SFI), a measure of motor function (0=Normal; −100=Nonfunctional) was −34 and −77 in MSC and vehicle groups, respectively (n≥9); with SNR it was −72 and −92 in MSC and vehicle groups, respectively. Long-term motor function (24 weeks) was apparent in MSC treated INR (SFI −63) but not in SNR (SFI −100) model. Gastrocnemius muscle atrophy was lower (P ≤0.05) in INR compared to SNR model. Significant reduction in axonal area (P ≤0.002), g ratio/axonal myelination (P ≤0.05), reduced nerve fiber density, and increased degenerating axons in the distal segment of the transected/repaired tibial nerve was observed compared to the contralateral naive tibial nerve. Histology and molecular studies are ongoing.

Conclusion

MSC therapy appears to promote peripheral nerve regeneration and functional recovery.
The Nrf2 activator dimethyl fumarate inhibits acute graft-versus-host disease with retention of graft-versus-tumor effects
Authors: Ma Shoubao , Gong Huanle , Han Jingjing , Liu Shuangzhu , Liu Haiyan , Wu Depei
Published: January 2023

Dimethyl fumarate (DMF), an activator of Nrf2, has been shown to have anti-inflammatory and immunomodulatory properties without significant immunosuppression. we therefore hypothesized that DMF could be a new candidate drug for the treatment of aGVHD with retention of GVT effect. Lethally irradiated BALB/c mice transplanted with B6 bone marrow plus splenocytes were administered DMF or vehicle solution by gavage daily from day -3 to day 3. DMF treatment inhibited aGVHD, accompanied by less weight loss and lower clinical score. DMF is capable of significantly inhibiting proliferation of alloreactive T cells in a dose-dependent manner in mixed lymphocyte reaction assay. Furthermore, the percentages of CD69+T cells and IFN-γ expressing T cells, were significantly increased in aGVHD target organs from DMF recipients. In addition, DMF treatment markedly increased the frequencies of Treg cells in target organs. Depletion of CD25+ cells in DMF recipients aggravated aGVHD mortality compared with IgG control recipients. Moreover, Treg polarizing assay suggested that DMF could promote Treg cell differentiation in vitro in a dose-dependent manner. In addition, No protective effects of DMF were observed in mice transplanted with BM and challenged with leukemia cells. However, DMF treatment significantly prolonged the survival in tumor-bearing mice that received an allogeneic BMT with donor splenocytes. we also observed that donor T cells recovered from DMF-treated recipients and controls showed comparable cytolytic activity against leukemia cells. In conclusion, our findings demonstrated that activation of Nrf2 with DMF may be of significant use in preventing aGVHD and preserving GVT responses.
Immunoregulatory function of kidney CD4−CD8− double negative (DN) αβ T cells is MHC dependent
Authors: Sadasivam Mohanraj , Noel Sanjeev , Lee Sul A , Rabb Hamid , Hamad Abdel Rahim A
Published: January 2023

Despite strong evidence for the role of immune cells in ischemia reperfusion injury (IRI) in transplant and native kidney, the underlying mechanisms are poorly understood. Furthermore, there is little data regarding unconventional T cells, usually present in small numbers in the kidney. We have recently shown that a subset of alpha/beta T cells that is double negative (DN) for both CD4 and CD8 coreceptors are present in significant numbers in normal kidney of mice and also present in human kidney biopsies. Unlike conventional CD4 and CD8 T cells, DN T cells are highly dividing in the steady and rapidly increases following IRI. In addition, kidney DN T cells secrete IL-10 and IL-27, possess an in vitro regulatory function and ameliorate AKI in mice. Unknown aspects of DN T cells included identification their MHC restriction elements and roles of various types of MHC haplotype in their homeostasis. Our most recent results show that DN T cells are heterogeneous as indicated by the partial effect of lack of β2m and MHC class II on their homeostasis. In addition, lack of β2m significantly reduce activated phenotype that is seen in kidney DN T cells. Conversely, reconstitution of β2m-deficient mice leads to activation and expansion of endogenous DN T cells, suggesting interactions between DN and CD8 T cells. Further dissection of kidney DN T cell biology will help us understand the pathogenesis of IRI and other immune mediated kidney diseases.
Studies on Antibodies. I
Authors: Banzhaf Edwin J. , Sugiura K. , Falk K. George
Published: January 2023

The antisera analyzed were the following:

Antidiphtheria globulin, 2900 units per cubic centimeter. Designated A in the tables.

Antidiphtheria globulin, 975 units per cubic centimeter. Designated H.

Antitetanus globulin, 1900 units per cubic centimeter. Designated B.

Antitetanus serum, 500 units per cubic centimeter. Designated F.

Antitetanus serum, 60 units per cubic centimeter. Designated G.

Antistreptococcus serum. Designated C.

Antigonoccocus serum. Designated I.

Antimeningococcus serum. Designated E.

Normal serum. Designated D.

The method of separating and concentrating the diphtheria and tetanus antitoxin together with the pseudoglobulin with which they are associated was carried out as follows:

The plasma from the immunized horse was diluted with an equal volume of water and heated to 61°C. An equal volume of saturated solution of ammonium sulphate was then added. The resulting precipitate contained the fibrinogen, euglobulin, pseudoglobulin and antitoxin. This precipitate was taken up with water and reprecipitated with an equal volume of saturated solution of ammonium sulphate to free it from traces of albumin.
Immunochemical Studies of the Proteins of Cat Hair
Authors: Wodehouse R. P.
Published: January 2023

Summary and Conclusions

Besides “keratin,” which makes up the greater part of the hair of most animals, there can be obtained from cat hair other proteins and these differ both in chemical characteristics and anaphylactogenic behavior from pure keratin and from a peptone which was produced by acid hydrolysis from pure keratin and which is called in these experiments “keratin peptone.”

Other proteins obtained from cat hair were as follows:

“Alkali metaprotein,” the protein dissolved from hair by weak alkali which is precipitated by neutralizing and faintly acidulating.

“Acid metaprotein,” the protein that is dissolved from hair by weak alkali and remains in solution after neutralizing and slightly acidulating but is precipitated by again making alkaline.

“Cat hair peptone,” the protein dissolved from the hair by weak alkali that remains in solution after the other two are precipitated as indicated above. It is obtained by evaporating the solution to dryness after removing the mineral salts by dialysis.

If these proteins dissolved from the hair by weak alkali are precipitated by other means their anaphylactogenic properties may be altered. For example:

“Protein preparation No. 2,” prepared by saturating the alkaline extract with ammonium sulphate; and

“Protein preparation No. 3,” precipitated from an alkaline extract (from which no protein could be obtained by neutralization or acidulation) by the addition of a small amount of MgCl2 were comparatively inactive.
Complement Fixation in Tuberculosis with the “Partial Antigens” of Deyke and Much
Authors: Woods Alan C. , Bushnell George E. , Maddux Cabell
Published: January 2023

Partial antigens have been prepared from the tubercle bacillus, by disintegrating the bacilli in 1 per cent lactic acid, filtering, and extracting with alcohol, at varying temperatures, and with ether. The resulting antigens appear to be composed chiefly of the various fatty acids, and neutral fat of the tubercle bacillus together with traces of protein.

One hundred and seventy-six normal and tuberculous sera were titrated against these partial antigens, and against whole bacillus emulsion antigen. Seventy-two of the sera obtained from clinically normal cases, and 91 per cent of the sera from cases classed clinically as inactive tuberculosis reacted negatively to these antigens. Ninety per cent of the sera from cases classed clinically as incipient tuberculosis, 87 per cent of the sera from cases classed as active tuberculosis, and 92 per cent of the sera from advanced tuberculous cases reacted positively to the partial antigens.

Sufficient syphilitic sera could not be obtained to make any determination of the proportion of syphilitic sera which would react positively to these antigens but judging from 27 positive Wassermanns obtained at the same time the sera were examined, there appears to be little relationship between the Wassermann reaction and the reaction to the partial antigens.
Complement Fixation in Acute Anterior Poliomyelitis
Authors: Kolmer John A. , Freese Anna E.
Published: January 2023

Complement fixation tests conducted with the cerebrospinal fluids of 130 persons suffering with acute anterior poliomyelitis and salt solution extracts of various tissues yielded from 6 to 16 per cent weakly positive reactions with extracts of spinal cord, pons and medulla, cerebellum, cerebrum and 15 to 17 per cent positive reactions with extracts of poliomyelitic liver and spleen. Control antigens of non-poliomyelitic human and rabbit liver yielded negative reactions, while extracts of spleen yielded from 5 to 7 per cent positive reactions. With alcoholic extracts of these tissues the tests were uniformly negative. The cerebrospinal fluids of normal persons and children suffering with whooping cough and measles yielded negative reactions with all extracts. The cerebrospinal fluids of several paretics yielded strongly positive reactions with all salt solution and alcoholic extracts. Complement fixation tests with the sera of 58 persons in the various stages of acute anterior poliomyelitis yielded slightly positive reactions with 2 to 4 per cent of sera with salt solution extracts of spinal cord, pons and medulla, cerebellum and cerebrum. Tests with extracts of poliomyelitic liver and spleen were uniformly negative, as were all tests employing alcoholic extracts of these different tissues. With polyvalent suspensions of diplococci and streptococci cultivated from persons suffering with acute anterior poliomyelitis, weakly positive reactions were observed with 6 to 9 per cent of cerebrospinal fluids of 35 persons suffering with the disease; with sera all tests were negative, as likewise all tests with fluids and sera employing suspensions of the diphtheroids and Gram-negative bacilli cultivated from poliomyelitis cases. These results indicate that the micrococci of poliomyelitis may produce antibodies during the course of the disease in a manner analogous to the production of antibodies by streptococci in scarlet fever. The Wassermann reaction in acute anterior poliomyelitis is uniformly negative with both cerebrospinal fluid and blood serum. These tests indicate that suitable salt solution extracts of various tissues from fatal cases of acute anterior poliomyelitis may serve in a very small percentage of cases to fix or absorb a small amount of complement in the presence of large amounts of cerebrospinal fluid, but the results in general are in accord with those reported by Wollstein, Gay and Lucas and the reactions are too irregular and weak to be of any practical value in the diagnosis of the disease.
The Relation Between Antigen and Antibody in the Living Animal
Authors: Weil Richard
Published: January 2023

It has been said that all sciences pass through three stages, that of philosophic anticipation, that of detailed observation, and that of careful generalization. The science of immunology is now in the second stage, and is engaged in gaining a mastery of the facts upon which reliable generalizations may in the future be based. Knowledge of the details of the science is increasing apace, and day by day the sure foundations are being laid of a permanent and well established theory.

Annually we meet to exchange with one another the fruits of the year's labors, and so to widen and deepen the stream of our knowledge. But, of necessity, we deal with sharply circumscribed problems, because we wish to give one another only such things as we have thoroughly proved and tested. This tacit, but general, law of these meetings was wisely broken last year in Dr. Jobling's presidential address, which aspired to give a broad and critical review of one very important aspect of immunological study.
The Vasomotor Depression in Canine Anaphylaxis
Authors: Weil Richard
Published: January 2023

In an earlier paper (1) it was shown that the coagulability of the blood in anaphylactic dogs is due directly to a reaction on the part of the cells of the liver. At the same time the suggestion was offered that the vasomotor depression night be due to a local congestive reaction of the liver, leading to a depletion of the general circulation, and in consequence, to a marked fall in blood pressure. This view was based on the observation that in very acute shock in dogs the liver alone of the abdominal organs presents an intense grade of congestion. The following experiments are in support of that suggestion.

A dog is sensitized actively by means of the intravenous injection of horse serum. After an interval of three weeks the animal is etherized and the abdomen is opened by a median epigastric incision.
A Plea for the Standardization of Reports of Agglutination Tests
Authors: Hadley Philip
Published: January 2023

Possibly the time has not yet arrived when it is desirable to standardize the methods employed in making the macroscopic agglutination test, although there is an increasing tendency upon the part of bacteriologists to employ the technique presented by Kolmer (1915) in which the dilutions of the serum to be tested start at 1:20 and proceed by doubling to give 1/40 1/80, 1/160, 1/320, etc.

But in the manner in which tests are reported for publication there is observable a great discrepancy on the part of different authors. For the sake of comparative work it would seem to be of advantage to possess a standardized method of reporting these results, whatever the technique of making the tests may have been.

A review of published results of agglutination tests indicates readily enough that the plus sign (+), for indicating a positive test, and the minus sign (-) or a zero (0), for indicating a negative result, have been commonly adopted.
Tissue Transplantation and Anaphylaxis
Authors: Loeb Leo
Published: January 2023

The results of various investigations in the field of anaphylaxis, especially those of Schultz, Dale, Weil and Manwaring, definitely point to the conclusion that in the process of sensitization and anaphylactic reaction the union of the antigen with various tissues, the elaboration of antibodies on the part of the cells to which the antigen has been anchored, and the additional union of circulating antigen with the sessile cell-antibodies, are, if not the only factors, at least very important ones. Under certain conditions these changes in the sensitized cells lead to functional reactions on the part of the cells that are responsible for the anaphylactic shock and other anaphylactic phenomena of a more or less acute character. But we know that under certain conditions there may be anaphylactic reactions that entail morphological changes. Arthus has shown that in a sensitized rabbit necrosis of the sensitized tissue may follow the subcutaneous injection of the antigen.
Loss of Immune Substances from the Body
Authors: Neill James M. , Gaspari Emidio L. , Mosley Robert A.
Published: January 2023

The paper reported a study of the elimination of “natural” diphtheria antitoxin in human urine. The urines were divided into three classes: those containing no detectable serum protein as obtained from persons whose renal permeability to protein was normal; those containing moderate amounts of protein as obtained from persons (cardiac failure and orthostatic albuminuria) of abnormal renal permeability but with no lesions in the lower urinary tract; those obtained from persons with lesions in the lower urinary tract that might have afforded opportunity for whole serum to leak into the urine without filtration through the kidney. When derived from persons with normal kidney permeability, detectable amounts of antitoxin (0.002 to 0.004 unit per 1.0 cc.) were found only in the urines voided by a man possessing an exceptionally high (15.0 units) concentration in his blood stream. When the urines contained moderate amounts of serum protein, antitoxin was contained if the donor of the urine possessed only moderately high amounts in his blood. The occurrence of antitoxin in urine containing no detectable serum protein is discussed.

Data on the daily output of antitoxin showed that elimination by way of the urine can account for only a small portion of the drop frequently observed in the antitoxin content of the blood. However, none of the pathological urines examined contained more than moderate amounts of albumin, and it is suggested that in cases of acute nephrosis or other clinical conditions in which large amounts of serum protein are eliminated, urinary elimination of antitoxin might be sufficient to cause a significant loss in the immunity reserves of the body.
A Study of the Acid Precipitation of Diphtheria Toxin
Authors: Bunney William E. , Cianciarulo J. , Kiamil Mumtaz
Published: January 2023

A study has been made of yields of diphtheria toxin obtained by acid precipitation of the crude toxin. A comparison has been made between the yields obtained when the acid precipitate is centrifuged immediately after acidification and when twenty-four hours storage in the cold intervenes between the acidification and the centrifugation. Results are obtained which indicate that a satisfactory method for large scale purification of diphtheria toxin would consist in the addition of acid to the point of maximum precipitation followed by immediate centrifugation, washing and solution of the precipitate at the desired concentration.
Studies on the Bacterial Toxins
Authors: Wadsworth Augustus , Quigley James J.
Published: January 2023

In recent years many attempts have been made to develop a simple and trustworthy method of concentrating and purifying the toxic substances in sterile filtrates of cultures of streptococci. Various methods have been followed but in general they consist of precipitating the protein-toxin fraction and then purifying by dialysis or ultrafiltration. Huntoon (1) used ammonium sulfate and acetic acid. Henry and Lewis (2) precipitated the toxin with alcohol. Hartley (3) dialyzed through collodion and then precipitated with acetic acid. Pulvertaft (4) precipitated whole broth culture with alcohol. Ando (5) precipitated his toxin with absolute alcohol. Shinn (6) claims to have secured the best results with acid alcohol. It is evident from the results obtained with these different methods that the toxic substances of the streptococcus can be concentrated and, to a certain extent, purified in the process.
Xanthone Jacareubin inhibits FcɛRI-dependent degranulation in mast cells by the blockage of extracellular calcium entry
Authors: Castillo-Arellano Jorge Ivan , Guzmán-Gutiérrez Silvia Laura , Ibarra-Sánchez Alfredo , Hernández-Ortega Simón , Reyes-Chilpa Ricardo , Gonzalez-Espinosa Claudia
Published: January 2023

Mast cells are important effectors in allergic reactions since they produce a number of pre-formed and de novo synthesized inflammatory compounds in response to high affinity IgE receptor (FcɛRI) crosslinking. IgE/Antigen-dependent degranulation and cytokine synthesis of this cell type have been recognized as relevant pharmacological targets for the control of deleterious inflammatory reactions. Despite the relevance of allergic diseases worldwide, pharmacological control of mast cell degranulation has not been achieved.

In this work we analyzed the effect of the natural xanthone jacareubin on the main activation parameters of this cell type utilizing bone marrow-derived mast cells (BMMCs).

Jacareubin, the xanthone obtained from the heartwood of the tropical tree Callophyllum brasilense, showed no toxicity when tested on bone marrow-derived mast cells (BMMCs) and did not activate degranulation by itself. However, jacareubin inhibited IgE/Ag-induced degranulation (determined as the secretion of β-hexosaminidase) in a dose-response manner (IC50 = 46 nM). Jacareubin blocked FcɛRI-dependent reactive oxygen species (ROS) formation, intracellular calcium mobilization and calcium-dependent protein kinase C (PKC) phosphorylation.

Detailed analysis suggest that at least one of the molecular actions of jacareubin relies on the inhibition of plasma membrane calcium channels involved in degranulation of mast cells. It is proposed to jacareubin as a natural drug with antioxidant activity and a regulator of calcium channels, with prophylactic potential for the treatment of allergies produced by mast cells.
Epigenetic regulation of mast cell function via inhibition of histone acetylation results in suppression of mast cell responses
Authors: Mathias Clinton B. , Krajewski Dylan , Kaczenski Edwin , Rovatti Jeffrey , Polukort Stephanie , Dollard Catherine , Mire Mohamed , Kinney Shannon RM.
Published: January 2023

Mast cells are highly versatile cells that perform a variety of functions depending on the immune trigger, context of activation and cytokine stimulus. Antigen-mediated mast cell responses are regulated by transcriptional processes that result in the induction of numerous genes contributing to mast cell function. Recently, we also showed that exposure to dietary agents with known epigenetic actions such as curcumin can suppress mast cell-mediated food allergy, suggesting that mast cell responses in vivo may be epigenetically regulated. To further assess the effects of epigenetic modifications on mast cell function, we examined the behavior of bone marrow-derived mast cells (BMMCs) in response to trichostatin A (TSA) treatment, a well-studied histone deacetylase inhibitor. IgE-mediated BMMC activation resulted in enhanced expression and secretion of IL-4, IL-6, TNF-a and IL-13. In contrast, pretreatment with either curcumin or TSA resulted in altered cytokine secretion. This was accompanied by decreased expression of FceRI and mast cell degranulation. Interestingly, exposure to non-IgE stimuli such as IL-33, was also affected by TSA treatment. Furthermore, long-term TSA exposure contributed to mast cell apoptosis and a decrease in survival. Further examination revealed a decrease in phospho-relA levels in TSA-treated BMMCs, suggesting that TSA alters transcriptional processes, resulting in decreased NF-kB activation. These data therefore suggest that the epigenetic regulation of mast cell activation during immune responses may occur via regulation of histone acetylation, and that exposure to dietary substances such as curcumin may induce epigenetic modifications that modulate mast cell function.
Diffuse panbronchiolitis in a patient with humoral immunodeficiency successfully treated with erythromycin
Authors: Saridakis Stephanie , Sandhu Monica , Jhaveri Devi K. , Hostoffer Robert W. , Matta Maroun , Tcheurekdjian Haig
Published: January 2023

Diffuse panbronchiolitis (DPB) is a rare and under diagnosed disease of the respiratory tract that affects the lungs bilaterally in addition to the layers of the respiratory bronchioles. The condition commonly occurs in patients of Japanese ancestry. However, as international migration increases, more cases are arising in the United States. Clinicians rarely consider DPB in the differential when evaluating patients with lung disease as DPB mimics several other pulmonary disorders, especially in patients with humoral immunodeficiencies. We report the first case of DPB in a patient with humoral immunodeficiency that was successfully treated with erythromycin as evidenced by symptom control and follow-up. As our understanding of humoral immunodeficiencies improves, so must the recognition of rare pulmonary diseases such as DPB, which can mimic more common pulmonary disorders seen with humoral immunodeficiencies. Treatment for DPB involves macrolide antibiotics such as erythromycin for a minimum of 6 months duration. We have concluded that DPB can be successfully treated in humoral immunodeficiency with macrolides.