bluuebunny/crossref_metadata_2025 huggingface dataset:
Outcome of Chlamydia muridarum genital infection and immune response in a beta2-adrenergic receptor knockout stress mouse model
Authors: Belay Tesfaye , Martin Elisha
Published: January 2023

The beta-2-adrenergic receptor (β2-AR) is one of the β-AR subsets found in most immune cells and is stimulated by norepinephrine (NE) binding. We showed previously that exposure of a mouse model to cold water stress leads to high level production of NE that may result in increased intensity of Chlamydia muridarum genital infection but the mechanism is not well defined. The purpose of this study was to develop and characterize a β2-AR knockout (KO) stress mouse model to investigate its susceptibility to C. muridarum genital infection. We hypothesized that lacking the β2-AR leads to a decreased C. muridarum shedding from the genital tract of stressed mice compared to that of stressed wild type (WT) mice. Stressed and non-stressed C57BL/6J WT and β2-AR KO mice were infected intravaginally with C. muridarum. Swabbing at 3-day intervals during primary infection and C. muridarum isolation was performed by culturing in McCoy tissue culture following standard methods. β2-AR KO stressed mice had a mean of 8540 inclusion forming units/ml (IFU/ml) compared to a mean of 18000 IFU/ml of stressed wild type at day 9 post infection. The overall course of primary infection in the β2-AR KO mouse model showed slightly reduced Chlamydia shedding. In addition, transfer of CD4+ T cells from black J6 WT to β2-AR KO resulted in a significant increase of shedding compared to that of non-stressed WT transfer (6000 versus 1000 IFU/ml). Secretion of interferon gamma was restored in β2-AR KO mice compared to WT. Overall, these results imply that deficiency in β2-AR leads to decreased C. muridarum shedding compared to the WT. However, other members of the alpha- and beta-adrenergic receptor family may compensate for the absence in β2-AR KO mice thus more investigation is needed.
Expediting antibody discovery using Bioelectronica’s HypercellTM platform
Authors: Yu Tian , Hull Jonathan , Ruiz Andrea , Bhat Ashwini , Basu Amar
Published: January 2023

Antibody-based drugs have been successful in a range of therapeutic categories. However, generating monoclonal antibodies is time-consuming and expensive. A common approach is Hybridoma technology, which overcomes the short life-span of IgG-secreting plasma B cells in vitro. However, many plasma B cells are lost due to the low efficiency of hybridoma cell fusion (typically <10%).

Direct single B cell screening strategies have emerged to bypass hybridoma fusion and recombinatorial display, coupled with the generation of recombinant monoclonal antibodies through mammalian expression systems. Obtaining expression systems with the required productivity, specificity and stability for clinical or commercial use requires screening millions of cells and thousands of clones.

Bioelectronica’s HypercellTM platform is an emerging technology used throughout antibody discovery and cell-line development to identify and isolate single, high-antibody secreting cells from large pools (~10,000,000 cells) in short periods (ca. 48 hrs). This scalable “electrofluidic” sorting system reduces time and cost by integrating antigen-detection reagents and real-time computer vision analysis to expedite single cell sorting.

In this paper antigen-specific IgG-secreting hybridoma cells are identified and sorted by their secretion rate. The cells and reagents are encapsulated in a Polydisperse Oblate Dispersion system (PODs), incubated for 1–4 hours for signal gain, and loaded into the HypercellTM device for cell sorting. Alternatively, the mixture can be analyzed without sorting to produce single cell secretion “finger print” signatures that can help identify unique expression patterns and monitor cell line stability over culturing time.
Development of a Chinese Herbal Immunogen Assay for the Measurement of T cell Anti-Tumor Immunity
Authors: Yen Hung-Rong , Song Ying-Chyi
Published: January 2023

T cells are the master regulators of adaptive immune responses. Proper T cell responses are essential for the hosts to orchestrate sufficient adaptive immunity. T cell immunity is crucial for anti-tumor immunity. Manipulating T cell immunity by Chinese herbs is a promising field to explore. Many herbs were reported to regulate the T cell activation. Modulation of T cell immunity by Chinese herbs may be an applicable intervention to modulate immunity, for example, to promote anti-tumor immunity against cancer. In this study, we developed a Chinese Herbal Immunogen Assay to measure the T cell immunity by screening the commonly used Chinese herbal formulas and single herbs from our Chinese herbal medicine extracts database. In the preliminary test, we collected human peripheral blood mononuclear cells (PBMC) were from fresh whole heparinized blood of healthy donors or cancer patients to proof the concept. The isolated PBMCs were stimulated with DMSO or a panel of various herbal extracts, respectively. Cytokines secretion from the activated PBMCs in culture supernatants were quantified at various time points by using standard ELISA for TNF-a, IL-6 and IFN-γ. At baseline, PBMCs from cancer patients had a lower average level of TNF-a, IL-6 and IFN-γ, compared to the healthy controls. After stimulation with a panel of herbal extracts, we identified the herbs that can consistently enhance cytokine productions. These herbs could be used for further development of new drug for boosting anti-tumor immunity.
Immunosenescent characteristics of T cells in young patients following haploidentical stem cell transplantation from parental donors
Authors: Lee Ga Hye , Hong Kyung Taek , Kim Bonah , Kang Yeon Jun , Lee Su Jeong , Choi Jung Yoon , Shin Hee Young , Lee Won-Woo , Kang Hyoung Jin
Published: January 2023

Pediatric and adolescent patients in need of allogeneic hematopoietic stem cell transplantation (HSCT) generally receive stem cells from older, unrelated or parental donors when a sibling donor is not available. Despite encouraging clinical outcomes, it has been suggested that immune reconstitution accompanied by increased replicative stress and a large difference between donor and recipient age may worsen immunosenescence in pediatric recipients. Therefore, in this study paired samples were collected at the same time from donors and recipients of haploidentical hematopoietic stem cell transplantation (HaploSCT). We then conducted flow cytometry-based phenotypic and functional analyses and telomere length (TL) measurements of 21 paired T-cell sets from parental donors and children who received T cell-replete HaploSCT with post-transplant cyclophosphamide (PTCy). Senescent T cells, CD28− or CD57+ cells, were significantly expanded in patients. Further, not only CD4+CD28− T cells, but also CD4+CD28+ T cells showed reduced cytokine production capacity and impaired polyfunctionality compared with parental donors, whereas their TCR-mediated proliferation capacity was comparable. Of note, the TL in patient T cells was preserved, or even slightly longer, in senescent T cells compared with donor cells. Regression analysis showed that senescent features of CD4+ and CD8+ T cells in patients were influenced by donor age and the frequency of CD28− cells, respectively. Our data suggest that in pediatric HaploSCT, premature immunosenescent changes occur in T cells from parental donors and therefore, long-term immune monitoring should be conducted.
Vesicular stomatitis virus expressing destabilized tumor associated antigens with enhanced MHC class I presentation to improve anti-tumor T cell responses
Authors: Huff Amanda , Evgin Laura , Thompson Jill , Kottke Tim , Wongthida Phonphimon , Driscoll Christopher , Shim Kevin G. , Scheulke Matthew , Vile Richard G.
Published: January 2023

We have previously shown that vesicular stomatitis virus (VSV) vectors encoding truncated tumor antigens, NRAS, Cytochrome C1, and TYRP1, prime superior systemic anti-tumor T cell responses and provide enhanced therapeutic benefit compared to vectors expressing full-length forms of the protein. Mapping of the roughly 20 amino C-terminal truncated regions onto structural domains of each protein suggested the truncations disrupted the stability and localization of each protein. Indeed, immunofluorescent staining and radiolabeled pulse chase found that truncated TYRP1 (ΔTYRP1) had an altered localization pattern and a significantly reduced half-life compared to full-length TYRP1. Thus, we hypothesized that expression of destabilized antigens from VSV would provide superior therapy compared to expression of wild-type antigens. To test this, we introduced point-mutations into structurally important domains of the model antigen ovalbumin (OVA). We identified one mutant which was significantly destabilized (OVAmut), which resulted in enhanced MHC Class I presentation of the immunogenic SIINFEKL epitope on transfected cell in vitro. Expression of OVAmut resulted in better recognition by cognate CD8 OTI T cells, enhanced T cell activation, proliferation, and effector cytokine production. Finally, a VSV vector expressing OVmut was better able to prime OTI T cells in vitro as well as in vivo. Ongoing studies aim to identify the changes in the quantity and repertoire of immunogenic epitopes presented from cells infected with VSV vectors expressing destabilized antigens. These studies provide insight into the capacity of the VSV platform to successfully encode unstable antigens with the goal improve immunogenic responses.
Tumor membrane vesicle-based vaccine immunotherapy inhibits tumor growth and metastasis of squamous cell carcinoma of head and neck in mice
Authors: Bommireddy Ramireddy , Munoz Luis E , Pack Chrstopher , Ramachandiran Sampath , Reddy Shaker , Chen Georgia , Saba Nabil , Shin Dong M , Selvaraj Periasamy
Published: January 2023

Head and neck cancer is a leading cause of cancer related deaths accounting for approximately 3% of all cancer related mortalities in the US. Currently, there is no cure for the advanced squamous cell carcinoma of the head and neck (SCCHN) thus development of efficacious therapies is urgently needed. To test whether vaccine-induced immunity inhibits tumor growth, we investigated efficacy of a tumor membrane-based vaccine (TMV) immunotherapy in murine SCCHN models. The MOC1, MOC2 and SCC VII tumors grown subcutaneously in syngeneic mice were harvested to generate TMVs. TMVs were then protein transferred with glycolipid-anchored immunostimulatory molecules GPI-B7.1 and GPI-IL-12 to generate the TMV vaccine. Mice were vaccinated with TMV vaccine after tumor cell challenge (therapeutic) and tumor growth was monitored every 3 days. Survival was then assessed using a Kaplan-Meier survival curve and significance determined using a Log-rank test for comparison analysis. The TMV vaccine inhibited tumor growth and improved the survival of mice challenged with MOC1 or SCC VII tumor cells. Interestingly, combination of anti-PD1 antibody with TMV vaccine decreased MOC1 tumor growth. However, MOC2 (poorly immunogenic tumor) did not respond to anti-PD1 or anti-CTLA-4 antibody therapy but significantly inhibited by TMV vaccine. MOC2 cells metastasize to the lungs in control mice but it is significantly inhibited in mice administered with TMV vaccine. TMV vaccine increased the infiltration of CD4+ and CD8+ T cells into the tumors suggesting that TMV vaccine converts cold tumors into hot tumors. These observations suggest that TMV vaccines can be harnessed to develop an effective immunotherapy for squamous cell carcinoma of the head and neck.
Characterization of innate and adaptive immune impacts of chemically dissimilar STING adjuvants
Authors: Boehm Dylan T , Mizuno Nobuyo , DeFilippis Victor
Published: January 2023

STING is a PRR for cyclic dinucleotides (CDN) synthesized by the cytosolic enzyme cGAS following exposure to microbial or cellular DNA. This stimulates innate responses characterized by localized type I IFN and proinflammatory cytokines. It is thus key to establishing adaptive immunity by facilitating APC recruitment and function. As such, STING is an attractive target to direct adjuvants and anti-tumor drugs. However, STING also drives processes such as apoptosis and autophagy that are certain to shape immune responsiveness. The degrees to which the protein elicits phenotypes can also vary substantially between STING-inducing stimuli including CDN species and small molecule agonists. While pharmacologic STING activation enhances immunogenicity of vaccine antigens, the impact of differences in STING-associated intracellular responses on overall immunity has not been explored. To address this, we utilize a mouse model of vaccination against Chikungunya virus. This is done in combination with chemically diverse STING agonists including CDNs and small molecules. Despite the direct engagement of STING by each, the characteristics of the immune reactivity to the antigen varied significantly. In addition, our exploration of STING-mediated innate correlates of these differences led to unexpected observations regarding agonist-specific cytokine induction, APC maturation, antigen trafficking and presentation. This has important implications in at least two areas: 1) It is crucial to filling the numerous gaps that exist with regard to understanding mechanisms that drive STING-associated immune enhancement; 2) It will reveal properties desirable for STING-based adjuvants to help direct future optimization.
Evaluation of Cancer-testis Antigens in Osteosarcoma and Dedifferentiated Liposarcoma as Targets for Immunotherapy
Authors: Jirovec Anna , Flaman Ashley , Purgina Bibianna , Werier Joel , Diallo Jean-Simon
Published: January 2023

T cell based immunotherapies targeting tumor antigens are a promising alternative to traditional cancer treatments due to their ability to eliminate malignant cells, but require highly immunogenic tumor-associated proteins as targets for CD8+ T cell recognition. Additional parameters to consider in a T cell based immunotherapy include tumor T cell infiltration and expression of HLA-peptide complexes on the surface of cancer cells. The goal of this study is to identify targetable immunogenic tumor antigens and to characterize the immune profile of human dedifferentiated liposarcoma (DDLPS) and osteosarcoma (OS) sarcoma subtypes by screening for cancer-testis antigen (CTA) expression, HLA expression, and tumor T cell infiltration by immunohistochemistry (IHC).

Human tissue micro-arrays composed of 80 cores of OS and 49 cores of DDLPS were obtained and stained by IHC for selected CTAs (MAGE-A3, NY-ESO-1 and SSX2) and various immune markers. In DDLPS, 11% of samples express MAGE-A3 and 2% express SSX2 and NY-ESO-1 in low percentages of tumor cells. In OS, 100% of samples express MAGE-A3 and 89% express SSX2, both with >80% of positive cases showing moderate to high expression. NY-ESO-1 is expressed in 78% of OS samples, predominantly at low levels. Brisk infiltration of CD8+ T cells is observed in over 70% of both sarcoma types. Furthermore, all sarcoma samples tested are positive for HLA expression.

These results show promising expression of CTAs MAGE-A3 and SSX2 in OS, which may be used as targets for immunotherapy for OS. The data generated throughout this study will provide insight into the immune profile of these sarcomas - information that is critical for immunotherapy design.
Dietary β-glucan reduced Salmonella shedding, shifted intestinal microbiome, and altered intestinal integrity
Authors: Byrne Kristen A , Trachsel Julian M , Bond Zahra F , Slate Jamison R , Kerr Brian J , Bearson Bradley L , Bearson Shawn M , Loving Crystal L
Published: January 2023

β-glucan is a prebiotic dietary fiber with immune and microbiota modulating properties. Dietary β-glucan can shift intestinal microbial populations and concentrations of short chain fatty acids (notably, butyrate and propionate), which may alter local populations of regulatory T-cells. Additionally, epigenetic reprograming of monocytes by β-glucan impacts subsequent monocyte responses to heterologous agonists, a form of innate training. Here, β-glucan from Saccharomyces cerevisiae was evaluated as a non-antibiotic dietary additive to limit shedding of the foodborne pathogen, Salmonella enterica server Typhimurium, via changes to the intestinal microbiota, intestinal integrity, and/or the local and peripheral immune system in pigs. Overall Salmonella shedding and cecal mucosa burden was significantly reduced in pigs fed dietary β-glucan compared to Salmonella challenged pigs on control diet. Shifts in microbial communities were associated with reduced Salmonella burden, as pigs fed β-glucan exhibited enhanced cecal epithelial barrier function, altered fecal bacterial communities, and increased short chain fatty acid concentrations. However, no significant differences in regulatory T cell population nor ex vivo monocyte responses were found between groups. Collectively, the non-antibiotic dietary additive β-glucan altered microbial communities and host epithelial cell function in conjunction with reduced Salmonella shedding in pigs.
Intraepithelial T cell phenotype and function diverge by intestinal location with age
Authors: Wiarda Jayne E , Trachsel Julian M , Bond Zahra F , Byrne Kristen A , Gabler Nicholas K , Loving Crystal L
Published: January 2023

Intestinal intraepithelial T cells (IETs) provide a first line of defense against intestinal pathogens and play an important role in immune tolerance versus activation. In pigs, intestinal T cell populations stabilize at ~8 weeks of age, but IET phenotypes and function are poorly understood during the early postnatal period. Better understanding age- and location-dependent characteristics of porcine IETs may be useful in developing strategies to maintain intestinal integrity and improve health. To assess impacts of age and intestinal location on abundance and composition of IETs, sections of jejunum, ileum, cecum, and colon from 4-, 6-, and 8-week-old pigs were analyzed by histologic staining and flow cytometric analyses. IET abundance increased with age, and CD2+CD8α+γδ and CD4−CD8α+αβ T cells were the primary IET populationsin all samples (>78%). Compositions of IETs were similar at all locations in 4-week-old pigs. As pigs aged, cecal and colonic IET compositions remained similar to each other but diverged from small intestinal IETs, while jejunal and ileal IET compositions diverged from both large intestinal IETs and each other. These findings indicate both intestinal location and age are drivers of IET heterogeneity. Small intestinal IETs had lower percentages of γδ IETs in 6- and 8-week-old pigs, and CD4−CD8α+αβ IETs increased in abundance at all intestinal locations as pigs aged. Percentages of CD27+IETs in ileum and large intestine decreased over time, but decreases were more drastic in large intestine, suggesting IETs in the distal but not proximal intestinal tract were activated as pigs aged. Small intestinal IET compositions closely resembled human IET populations, highlighting pigs as a biomedical model for IET research.
Immune fitness as a measure of animal health, welfare and productivity in the feedlot
Authors: Purdie Auriol C , Plain Karren , de Silva Kumudika
Published: January 2023

Background

Animals have long been bred for production traits that enable high-yield, but this does not consider the overall immunological ‘fitness’. What is the physiological basis that enables some animals to thrive or fail when exposed to production stresses, are there any measures of the immune component to identify these animals? Further, are there commonly related genes associated with immune fitness?

Methods

50 mixed breed beef cattle were randomly selected at feedlot intake. Blood and saliva samples were collected at arrival (high stress), post- acclimatization and pre-slaughter (60 days post intake or ≥400kg). Body condition and other measures (weight, height, disposition, coat colour) were collected at each sampling and at slaughter. Samples were processed for cell counts, RNA (transcriptomics analysis) and serum (protein analysis). Transcriptomic and proteomic analysis was by 24k Bovine Affymetrix ST1 GeneChip and RayBiotech Bovine Cytokine Array Q30.

Results and conclusion

Analysis correlated to outcome measures identified genes and proteins associated with immune fitness including those correlated to animals with susceptibility to disease and/or low weight gains. These results suggest that immune measures may be utilized to identify animals at risk of poor production performance facilitating improved animal management to optimize animal health and welfare.
TARGETING METABOLIC REPROGRAMMING OF RESPIRATORY SYNCYTIAL VIRUS INFECTION
Authors: Bahadoran Azadeh , Bezavada Lavanya , Rezinciuc Svetlana , Devincenzo John , Cormier Stephania , Smallwood Heather
Published: January 2023

Respiratory syncytial virus (RSV) causes severe respiratory disease and infects virtually most children by 2 years of age. It is the leading cause of hospitalization of infants worldwide and remerges later in life to be a serious lower respiratory tract illness in the elderly. Recently, significant emphasis focused on changes in host cellular metabolism in response to viral infection. We performed a retrospective study of pediatric patients infected with RSV and found this infection correlated with increased glucose uptake in the lungs. We characterized cells freshly isolated from naturally RSV infected pediatric patients’ nasopharyngeal aspirates (NPA) and quantified their bioenergetics. Next, we will determine RSV infection induced changes in metabolic responses of different cell types including primary human epithelial cells (NHBE), dendritic cells (DCs), macrophages (MOs) and A549 cell line. Additionally, we will characterize the infect co-cultures of NHBE and DCs/MOs with RSV to determine if their crosstalk can help regulate aberrant NHBE metabolism. We found a dramatic increase in glycolysis and mitochondria respiration in cells isolated from patients’ NPA. RSV infection increases basal respiration, ATP production, and proton leak while reducing mitochondrial oxygen consumption and respiratory capacity in pediatric patients’ NPA. Significant increases in metabolism in the infected patient’s upper respiratory cells ex vivo support our in situ findings in RSV infected patients lungs. Understanding and defining the metabolic changes in the host during RSV infection may lead to novel therapeutic approaches through targeted inhibition of specific cellular metabolic pathways.
Influenza virus suppresses the adaptive immune response leaving immune hosts vulnerable to influenza and other respiratory pathogens
Authors: Bohannon Caitlin D , Ende Zachary , Cao Weiping , Mboko Wadzanai , Ranjan Priya , Kumar Amrita , Mishina Margarita , Amoah Samuel , Gangappa Shivaprakash , Mittal Suresh K , Garcia-Sastre Adolfo , Pfeifer Blaine , Davidson Bruce A , Knight Paul , Sambhara Suryaprakash
Published: January 2023

Influenza infections are associated with several million cases of severe respiratory illness, hospitalizations, and hundreds of thousands of deaths globally. Secondary infections further complicate influenza’s high morbidity and mortality, and significantly factored into the severity of the 1918, 1968, and 2009 pandemics. The most common coinfections are bacterial, leading to bacterial pneumonia, though viral secondary infections also occur. Previous studies have shown that influenza can target innate responses and damage affected tissues, allowing for secondary infections. In this study we show that influenza virus targets not only innate immune responses but also the adaptive responses - specifically activated B cells, T cells, and NKT cells. Importantly, we demonstrate that infection with influenza virus can attenuate the adaptive responses to prior influenza vaccination and to other respiratory pathogen vaccinations, in humans and in mouse models. This occurs through a viral hijacking of the normal immune responses by taking advantage of elevated expression of sialic acid receptors on activated lymphocytes to preferentially infect and kill immune responders. Our findings provide a novel potential mechanism for the high incidence of secondary respiratory infections due to bacteria and other viruses, as well as for vaccine failures against other infectious agents during influenza seasons.
Human influenza A-specific memory T cells reside within multiple tissues and elicit a tissue-specific antiviral response when presented with viral antigens
Authors: Poon Maya M. , Meng Wenzhao , Dogra Pranay , Lam Nora , Szabo Peter A , Grifoni Alba , Tian Yuan , Kubota Masaru , Matsumoto Rei , Sette Alessandro , Prak Eline T Luning , Farber Donna L
Published: January 2023

Generation of memory T cells specific for conserved viral epitopes provides heterosubtypic protection to influenza infection. Previous mouse studies have shown that tissue-resident memory T cells (TRM) are critical for optimal protection. However, knowledge of human antiviral response is largely derived from blood. With our unique human donor tissue resource established in collaboration with LiveOnNY, we investigate the maintenance of influenza-specific T cells across diverse tissue sites and their response when presented with viral antigens. Using multimer staining and flow cytometry, we show that influenza-specific CD8 T cells are maintained in the lung tissue site of infection, as well as in blood, bone marrow, multiple lymph nodes, and small intestines. Compared to T cells recognizing persistent cytomegalovirus infection, influenza-specific T cells display higher levels of canonical residency markers—CD69 and CD103. Additionally, deep sequencing of the T cell receptor CDR3b repertoire reveals that influenza-specific T cells isolated from human tissue donors without active infection are clonally expanded and display a high degree of clonal overlap across tissue sites. When presented with viral antigens, influenza-specific T cells in all sites are polyfunctional and rapidly respond through cytokine production. Together, these studies reveal the dynamics of immune memory maintenance and antiviral response across tissues in the human body in response to influenza A virus.
Studies on Crystalline Urease
Authors: Hotchkiss Margaret , Tauber Henry
Published: January 2023

Crystalline urease in solution produced agglutination of the erythrocytes of the rabbit and of the mouse, but not of the sheep.

In the tests where high concentrations of urease were used hemolysis occurred, complicating the results. This was subsequently controlled by the use of a buffer solution, when agglutination could be readily seen.

After agglutination the urease was found to be active.

Agglutination could not be demonstrated in the whole blood of rabbits dead after urease injection.

Crystalline urease represents a highly purified form of toxic hemagglutinin (10).
The Use of Ethyl Alcohol as Precipitant in the Concentration of Antipneumococcus Serum
Authors: Felton Lloyd D.
Published: January 2023

Summary and Conclusions

Studies of the solubility of the pneumococcus antibody in alcohol reveal the following:

Practically all the protective substance present in the original serum is insoluble in 15 to 20 per cent alcohol at 0°C. The precipitated protein containing both water-soluble and water-insoluble protein can be washed with water to remove the former, and the latter dissolved in salt solution is found to contain protective antibody in practically the same concentration as the original serum. Precipitation with 10 per cent alcohol gives a product in which the immune protein is approximately 80 per cent of the total protein. Irrespective of hydrogen ion concentration from pH 5 to pH 9, if the precipitate is neutralized before diluting, the pneumococcus antibody is insoluble in 20 per cent alcohol. In more alkaline solutions a greater concentration of alcohol is necessary to cause precipitation. Water-alcohol mixtures precipitate the antibody but with an increased amount of inert protein as compared to the precipitate obtained with alcohol mixed with undiluted serum. The use of alcohol as a precipitant in the concentration of anti-pneumococcus serum gives a yield of at least 80 per cent of the protective substance found in the original serum. It has been found possible to dry the active material with retention of immunological characteristics.
A Study of Action of Saliva and Extract of Tonsils Upon Diphtheria Bacillus and Diphtheria Toxin
Authors: Bézi István
Published: January 2023

Summary and Conclusions

This preliminary report records the results of experimental tests of the action of saliva and extracts of tonsils and other organs upon diphtheria bacilli and diphtheria toxin. Evidence is presented which seems to indicate that some specimens of saliva may exert an anti-bacterial action against diphtheria bacilli and that saline extracts of tonsils and lymph nodes may inhibit the necrotizing action of diphtheria toxin. Whether or not these reactions are specific, has not been determined and the experiments do not permit the drawing of definite conclusions at this time.
The Effect of Alum on Horses Used for the Production of Diphtheria Antitoxin
Authors: Leonard George F. , Varley James R.
Published: January 2023

Summary and Conclusions

Three groups of horses were immunized for the production of diphtheria antitoxin. Group 1 received toxoid containing 0.5 per cent potassium alum, followed by diphtheria toxin containing alum. Group 2 received diphtheria toxoid, followed by diphtheria toxin—both without alum. Group 3 received diphtheria toxin-antitoxin, followed by diphtheria toxin—both without alum. As a result of the marked intolerance of the horses of group 1 toward alum antigens, they received less than threetenths of the antigen dosage given to the horses of Group 2. At trial bleedings after three months the average antitoxin titers per cubic centimeter for the horses of groups 1 and 2 were practically the same, namely, 360 units and 350 units, respectively; while the average titer of group 3 was 580 units. Based on the relative antigen doses and the subsequent antitoxin titers, the antigenic effectiveness of immunization with alum-treated toxoid and toxin was about three times as great as that of toxoid and toxin without addition of alum. A cumulative toxic effect was observed in the horses of group 1 after repeated doses of alum combined with diphtheria antigens. During a period of several weeks after injections containing alum had been discontinued, most of the horses in this group gradually declined and two-thirds of them died with symptoms of alum poisoning. The disastrous late after-effects upon horses of repeated injections of alum along with diphtheria antigens completely overshadow any increase in immunizing power attributable to the addition of the alum. It appears improbable, however, that the addition of a small amount of alum to the two or three doses of toxoid used for human immunization would exert any harmful effect.
Serological Studies on Iodinated Sera
Authors: Jacobs John
Published: January 2023

When iodine is added to animal sera without the presence of acid or alkali, substances are formed which precipitate with antisera prepared from iodinated sera. This change takes place in the presence of aqueous solutions as dilute as n/32. Wormall's observation that diiodotyrosine, but not potassium iodide, inhibits precipitation specifically in systems of iodinated proteins and their antisera, was confirmed. The specificity of substances formed in sera iodinated in neutral solutions was tested in several ways. Their specificity is similar to that of other iodinated proteins.
Additional Note to the Article “Studies on Tobacco Hypersensitivity”
Authors: Sulzberger Marion B.
Published: January 2023

In my article entitled “Studies on Tobacco Hypersensitivity,” appearing in the January, 1933, issue of the Journal of Immunology, the third and fourth paragraphs refer to my suggestions to Dr. Silbert in June, 1931, concerning skin tests with tobacco in thromboangiitis obliterans, and to the experiments which were published about a year and a half later by Harkavy, Hebald and Silbert. My object was simply to report the facts concerning the chronological sequence of events.

I realize now that these paragraphs may be susceptible to mis-interpretation. I therefore state that, to the best of my knowledge, Dr. Harkavy may not have known of my previous conversation and arrangement with Dr. Silbert. I feel it only just to affirm that it was not my intention to suggest, either directly or by implication, that Dr. Harkavy, when he commenced his investigations at a later date, knew of my own or any one else's previous interest or ideas in this field, nor did I mean to imply that he did not subsequently conceive and undertake his research independently.
Inheritance as a Factor in Resistance to an Infectious Disease
Authors: Irwin M. R.
Published: January 2023

Genetic and immunological tests gave no indication of an influence of passive immunity in the experiments.

The progeny from either carrier or non-carrier parents tended to be resistant only if so determined by their line of descent.

There was no difference in the proportions of carriers within the surviving litters of carrier or non-carrier parents.
The Effect of Staphylococcus Bacteriophage Lysin on Resistant Strains of Staphylococci
Authors: Rakieten Morris L.
Published: January 2023

A large proportion of the investigations dealing with bacteriophage have been concerned with the isolation of an active lytic principle against many different strains of bacteria. These studies, in general, have varied but little from the classic experiments of d'Herelle dealing with the isolation and development of several true bacteriophages (1917). This phenomenon is now so well known to most students of bacteriology that there are relatively few laboratories where the reaction between a susceptible strain of bacteria and a potent bacteriophage has not been observed. Since the initial discovery was described an enormous literature has accumulated bearing upon the relationship between bacteria and bacteriophage. As early as 1920 d'Herelle had shown that this infravisible being is responsible for bringing about marked changes in bacteria. Subsequently, he, as well as many others, have studied the transformations of various bacterial species brought about through the agency of bacteriophage.
Studies on Tissue Reactions in Immunity
Authors: Kahn Reuben L.
Published: January 2023

The desensitization of rabbits sensitized to complex proteins, such as human serum, was considered in a previous article (1). It was shown that, after an intravenous injection of a moderate amount of specific protein in a sensitized animal, no skin sensitivity reactions are obtained and the serum precipitins apparently disappear from the circulation; also that this desensitization phase is of comparatively short duration, that within twenty-four hours after the intravenous injection the animal begins to show a tendency to return to the sensitized state. Early experiments dealing with specific desensitization of rabbits that are sensitized (or immunized) to different organisms led to several observations which were not identical to those made in connection with desensitization to proteins. It was noted, for example, that an intravenous desensitizing injection of a suspension of typhoid bacilli in rabbits sensitized to these organisms, caused a reduction in the skin sensitivity response, but produced no apparent effect on the serum agglutinins.
Is the Eberson Colloidal Gold Test Specific for Poliomyelitis Antibodies?
Authors: Schultz E. W. , Clifton C. E. , Gebhardt L. P. , Chambers J. V.
Published: January 2023

Summary and Conclusions

Colloidal gold tests carried out according to the procedure described by Eberson did not enable us to distinguish between serums collected from poliomyelitis convalescent monkeys and normal monkeys (M. rhesus). The test therefore does not seem to be in any sense specific for the detection of poliomyelitis antibodies. The reactions described are apparently due to an adsorption of gold particles by some unidentified ingredients of the serum with a subsequent “settling out” of these aggregates after becoming “coated” with gold. The substance in the serum which is responsible for this phenomenon seems to fluctuate in amount and may be closely related to food stuffs absorbed by the blood from the alimentary tract.
Studies on Vascularization of the Cornea
Authors: Julianelle L. A. , Morris M. C. , Harrison R. W.
Published: January 2023

Summary and Conclusions

Vascularization of the cornea of rabbits has been studied as a response to bacterial hypersensitiveness. The organisms utilized for inducing hypersensitiveness have been Staphylococcus aureus, Pneumococcus and Bact. granulosis. These bacteria have been administered repeatedly by intracutaneous and sub-conjunctival injections: By conjunctival instillations; a, without corneal scarification; b, with corneal scarification; c, with mixtures of broth cultures and sand; and d, by preliminary instillation of homologous broth filtrates a day preceding the administration of bacteria. Staphylococcus aureus has been the most consistent sensitizing antigen, and instillation with corneal scarification, the most effective method of inducing vascularization of the cornea. Direct corneal vascularization occurs only when the cornea is injured slightly but definitely, but similar trauma per se does not cause this reaction. The corneal vascularity observed during early development resembled pannus tenuis of trachoma, and that observed in the latter stages was suggestive of pannus crassus. It was not possible to correlate corneal sensitivity of the inoculated eye with the presence or absence of corneal sensitivity of the uninoculated eye or to heightened skin reactivity to the homologous bacterial nucleoprotein, or to the presence or absence of circulating antibodies. The antibody response following conjunctival instillations in its various forms was slight or entirely lacking. While the indications are that the vascularity described in this study is in response to bacterial hypersensitiveness, no experiments have been undertaken as yet to indicate whether pannus in trachoma is initiated by a similar stimulus.
Group Specificity of Dried Muscle and Saliva
Authors: Boyd William C. , Boyd Lyle G.
Published: January 2023

By differential adsorption both of human and of group-specific immune sera it was possible to demonstrate agglutinogens A and B in dried human muscle in exact conformity to the group previously determined in the erythrocytes. Agglutinogens M and N could not be demonstrated in dried muscle, or in dried or fresh saliva, though in conformity to the reports of other workers, A and B could be found in fresh or dried saliva.
The Effect of Reticulo-Endothelial Cell Blockade Upon Antibody Formation in Rabbits
Authors: Tuft Louis , Mulrooney Mary
Published: January 2023

The site of formation of many types of immune antibodies has for many years been a moot question among immunologists. By some it was thought that the blood forming cells of the bone marrow or spleen possessed this function; by others because of its phagocytic properties, the polymorphonuclear leucocyte. In more recent years with the acquisition of more knowledge concerning the cells of the reticulo-endothelial system, the property of antibody formation has been attributed to these cells and upon this question considerable discussion has arisen in recent literature. The difficulty in proving such an assumption lies in the fact that the reticulo-endothelial cells are widespread throughout many tissues and organs; hence their function cannot be studied experimentally by surgical removal in the same fashion as the function of such an organ as the spleen. One must resort instead to methods designed to suppress or blockade such functions.
Studies of Scarlet Fever
Authors: Hooker Sanford B. , Follensby Edna M.
Published: January 2023

Numerous reliably observed discrepancies to the single-toxin hypothesis of the mechanism of scarlet fever are readily explained by merely assuming that multiple antigenic toxins exist. This report demonstrates the existence of two such toxins, A and B, whose qualitative distinctiveness is evidenced both by immunologic and chemical criteria. These include specific antigenicity, neutralizability, and erythrogenic activity, the effects of dialysis, adsorbents, precipitants, and enzymic digestion. Other erythrogenic streptococcal toxins undoubtedly exist. A single toxigenic strain may produce A or B or both or neither.
The Prozone Phenomenon in Specific Bacterial Agglutination
Authors: Jones F. S. , Orcutt Marion
Published: January 2023

The results of experiments with two agglutination inhibitory sera specific for B. abortus indicate that when proper proportions of either are added to a strong B. abortus agglutinin, agglutination is inhibited or a prozone developed. Bacteria not agglutinated in the prozone serum can be centrifuged and resuspended in the same mixture and remain in suspension. When the original supernatant is replaced with salt solution agglutination usually occurs promptly although where the concentration of prozone serum is considerable an additional washing with sodium chloride may be required to induce clumping.

In respect to electrical charge and migration in an electrical field and increases in antigenic volume, B. abortus behaves similarly in both inhibitory and agglutinating sera. Differences in behavior between antigens sensitized to the two types of sera were brought out in the rate of sedimentation as the result of centrifugation and in the force required to separate films of B. abortus under both conditions.

The failure to agglutinate is attributed to the deposition on the surface of the deposited globulin film of a substance which interferes with the cohesive properties of specifically sensitized organisms.
On Immunization Against Typhoid Fever with Heterobacterigen
Authors: Suzuki Ch. , Sugio K.
Published: January 2023

Utilizing as antigens heat-killed cultures of Bact. pullorum and Bact. sanguinarium, which are probably not pathogenic for man, the authors have succeeded, by the oral method of Besredka, in provoking the production of antibodies against the pathogenic agents of typhoid fever, paratyphoid fever and bacillary dysentery (Shiga).

The authors wish to apply this result to the treatment and prophylaxis of these infectious diseases.

The preparation of the two bacteria has been named heterobacterigen for the sake of convenience.

The bacteriological prophylaxis and treatment hitherto applied to various bacillary diseases have been restricted to using preparations of the bacteria causing them, and this is regarded as theoretically the most reasonable measure.

It is the purpose of this paper to call attention to the possibility of establishing immunity against some bacterial diseases with bacteria other than those etiologically concerned in those diseases; and for convenience we shall designate such “other bacteria” as heterobacteria.
Hemolytic Streptococcus Toxins and Antitoxins
Authors: Malcolm Wilbur G. , Wyman Louise
Published: January 2023

The medium and method described here for the preparation of streptococcus toxin has been employed by us since October, 1931. The medium is a modification of the one used for some time for diphtheria toxin production. The streptococcus medium differs chiefly in that Pfanstiehl or Witte peptone has been substituted for Difco Proteose peptone, and it contains dextrose. The adjustment of the reaction during the growth period is considered essential.

Method.

Fresh veal (leg) infusion. One part meat in 2 parts water by weight. Care should be taken to have the meat as free as possible of fat and connective tissue. Refrigerate overnight. Place in double boiler—raise temperature very slowly to 80°C. (Should require between four and five hours.) Filter through one layer of cheesecloth. (Let drain, do not press.) Weigh and make up to the original weight of the water and meat, minus the weight of the cooked meat. Let stand five or ten minutes to settle, and then syphon off the supernatant.
Some Observations on the Relative Importance of the Reticulo-Endothelial Tissues and the Circulating Antibody in Immunity
Authors: Teale F. H.
Published: January 2023

The above experiments therefore show that with regard to: The highly virulent streptococcus. The immunised animal can completely clear the peripheral circulation and prevent the occurrence of secondary waves of bacteriaemia, in the absence of (1) agglutinin, (2) any germicidal power of the whole blood, and (3) any protective antibody capable of conferring passive immunity.B. anthracis. The same results as a.Pneumococcus type III. The immunised animal can clear the circulation and prevent the occurrence of secondary waves of bacteriaemia even though (1) the blood cannot confer passive protection on mice, (2) the blood is not germicidal (except occasionally, experiment 25), (3) the blood does not have any appreciable power of conferring passive protection on rabbits (when the results are compared with the effect of the inoculation of normal sera, etc., into normal rabbits), and (4) the serum contains no agglutinin nor precipitin against the organism.Animals immunised a long time previously with pneumococcus type I, where all traces of agglutinin had disappeared and where there was no germicidal or protective power to cause passive protection in mice or rabbits, completely cleared the circulation of the actively growing virulent organism.With the B. Friedländer, the same was noted in animals immunised long previously and which had completely lost their agglutinating antibody.

All these experiments taken together irresistably lead to the conclusion that the chief factor in immunity is the state of the tissues with regard to the infecting bacteria in question.
Agglutinins for Sheep and Rabbit Erythrocytes in Human Sera
Authors: Stuart C. A. , Tallman Juanita , Anderson E. G. E.
Published: January 2023

An apparent relationship has been shown between the titers for sheep and rabbit erythrocyte agglutinins in the blood of man. In the 760 human sera examined a relationship has been shown between the agglutinin titers for sheep cells at 37° and at 5°C. This applies also to rabbit cell agglutinins. A statistically significant sex difference was found in both sheep and rabbit agglutinin titers in human sera, with the higher agglutinin content in female sera.
Cutaneous Reactions and Antibody Response to Intracutaneous Injections of Pneumococcus Polysaccharides
Authors: Finland Maxwell , Dowling Harry F.
Published: January 2023

Immediate cutaneous reactions, similar to those described by Tillett and Francis (15), were elicited in patients recovering from pneumonia. These reactions were, in general, type-specific and were associated with the homologous type antibody in the serum. Very similar reactions were elicited with preparations of soluble specific substances and cellular carbohydrates prepared from the same pneumococcus types by the methods of Heidelberger, Goebel and Avery (18) and those of Wadsworth and Brown (10), respectively.

Characteristic delayed reactions were seen only with the cellular carbohydrates. These were not associated with type-specific antibodies. They were most frequently observed with the cellular carbohydrates of the atypical type I pneumococcus and with that obtained from the virulent type I strain.

Delayed cutaneous reactions with the cellular carbohydrate of an atypical type I pneumococcus were obtained regularly during the febrile stage of a variety of infectious disease and could not be elicited soon after recovery in such cases. These findings are similar to those obtained by Francis and Abernethy (17) with the “C” substance of Tillett, Goebel and Avery (19).

All of the type-specific polysaccharides were antigenically active in human subjects. They produced in almost every instance a strictly type-specific antibody response. Only minor differences were observed with the different preparations of types II and III polysaccharides. The type I cellular carbohydrate, however, was quantitatively more active than the corresponding SSS in this respect. Occasional subjects showed specific antibodies, particularly against type I pneumococci, following injections of the atypical cellular carbohydrate.