bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of immune cells of stressed beta2-adrenergic receptor homozygous knockout mouse model during Chlamydia muridarum genital infection
Authors: Belay Tesfaye
Published: January 2023

The relationship of stress in chlamydia genital disease pathogenesis and its influence on the host immune response remains unknown. Beta2-adrenergic receptor(β2-AR), the major receptor of the stress hormone norepinephrine (NE), is known to impair the function of immune cells. This study was undertaken to determine whether cold-induced stress (CIS) results in differential viability percent of immune cells β2-AR KO and WT C57BL/6J treated with NE, β2-AR agonist(Fenoterol), or antagonist(ICI 118,551), and patterns of cytokine production. CIS was induced by immersing mice in cold water for five minutes daily for 21 days. Total CD4+, CD8+, differentiated bone marrow-derived monocytes were purified by negative selection and treated with chemicals for one hour before proliferation for 48 h proliferation. The vi-cell viability analyzer showed that NE results in a 30 and 40 % decrease in viability of WT C57BL/6J CD4+ T and CD8+ cells, respectively, compared to KO (p<0.05). Similarly, NE and Fenoterol resulted in a 20 and 27% decrease in viability of bone marrow-derived monocytes compared to the β2-AR KO mice. Cells treated with ICI 118, 551 showed the slightest reduction in viability (12%) compared to those treated with NE. ELISA results show decreased IL-4 and increased IFN-g secretion by CD4+ T cells of β2-AR KO compared to WT. The secretion of TNF-a by CD8+ cells and bone marrow cells treated with NE and β2-AR agonist or antagonist was high in all treatment groups. This study provides insights for the first time to better understand the NE-β2-AR pathway during chlamydia genital infection.

Supported by WV-INBRE of NIH Grant P20GM103434 and NIH grant # 1R15AI124156-01 given to BSC.
Nonencapsulated Streptococcus pneumoniae cause Bacteremia through Inhibition of C-Reactive Protein Binding
Authors: McDaniel Larry S , Thompson Courtney , Keller Lance
Published: January 2023

Naturally occurring nonecapsulated Streptococcus pneumoniae (NESp) are found worldwide and can cause invasive infections. In contrast, encapsulated pneumococci that fail to express capsule are rapidly cleared from the blood of infected mice. We previously demonstrated that NESp expressing the Ami-like proteins AliC and AliD are able to evade complement-mediated killing. Data indicated that factor H (FH) failed to bind to NESp, and thus did not reduce complement activation as seen in FH binding pneumococci. Proteomic data have shown that deletion of aliC and aliD altered pneumococcal protein expression. Additionally, NESp expressing AliC and AliD caused invasive pneumococcal disease in C1q-deficient mice. The purpose of this study was to examine the regulated pneumococcal genes in complement evasion by NESp. Flow cytometry was used to examine binding of C3b to NESp. A mouse model of infection was used to investigate complement interaction with NESp in vivo. NESp expressing AliC and AliD were able to persist for at least four days following intravenous infection of mice. Deletion of aliC and aliD resulted in the clearance of NESp within 24 hours post-infection. There was a four-fold increase in C3b deposition on the mutant NESp compared to the wildtype. Choline binding protein AC (CbpAC), a variant of pneumococcal protein A (PspA), was found to have reduced expression in the aliC and aliD deleted mutant. C-reactive protein (CRP) showed increased binding to NESp in which cbpAC was deleted. Regulated expression of CbpAC in NESp is able to overcome the innate immune response and lead to persistence of NESp within the host by reducing C1q deposition and arresting classical complement activation.

Supported by Departmental research funds.
The diversity of beta defensins in lungfish (Dipnoi)
Authors: DeMmon Diego M , Benedicenti Ottavia , Casadei Elisa , Salinas Irene
Published: January 2023

Mucosal surfaces are the first line of defense against pathogens. At these surfaces, mucus containing innate and adaptive immune molecules is constantly secreted to protect the animal host. African lungfish are the extant relatives to all tetrapods and have a dual mode of living, swimming in freshwater during the wet seasons and aestivating during the dry seasons. During terrestrialization, African lungfish produce large amounts of mucus that form a dry cocoon to prevent water loss. We recently reported that the lungfish cocoon acts as a bacterial trapping structure and it is rich in antimicrobial peptides (AMPs), specifically beta defensins (BD). As many AMPs, BDs are small amphipathic molecules and mainly cationic. The goal of this study is to provide a full characterization of BD genes from transcriptomes and genomes of different lungfish species. Using the canonical six cysteine motif from vertebrate BD molecules, we searched for all BD genes Protopterus dolloi transcriptomes, the genomes of P. annectens and Australian lungfish (a non-aestivating lungfish, Neoceratodus forsteri). BLASTs searches identified 9 BD genes in P. dolloi transcriptomes with predicted highly cationic characteristics except for BD-7. Sequence alignment and phylogenetic trees indicate that three P. dolloi BD genes may be an intermediate form of AMP between beta and alpha defensins. Genomic analyses indicate that P. annectens BD genes contain 3 exons and 2 introns similar to all defensin genes. Future studies will determine the tissue distribution and antimicrobial function of Protopterus spp. BD molecules and determine their potential application as alternatives to antibiotics.

Supported by National Science Foundation Integral Organismal Systems (IOS) award # 1938816
Group B Streptococcus does not translocate the gut epithelium of neonatal mice.
Authors: Harlow Olivia S , Greenfield Kara G , Knoop Kathryn
Published: January 2023

Neonatal sepsis remains a leading cause of death in preterm and low-birthweight infants and can be separated into two groups: early-onset sepsis (EOS) and late-onset sepsis (LOS). Previous studies have shown that LOS can result from translocation of enteric bacteria across the intestinal epithelium, resulting in bloodstream infection. Through metagenomic sequencing we have observed, unsurprisingly, potential sepsis-causing organisms such as Escherichia coli, Staphylococcus aureus, Enterobacter cloacae, and Group B Streptococcus (GBS) in the stool of neonatal intensive care unit patients. Our previous work has shown E. coli can disseminate from the intestines via goblet cells in a model of LOS, but how other bacterial pathogens disseminate remains unclear. GBS is often associated with EOS following in utero infections and vertical transmission from the mother during pregnancy and delivery. However, the ability of gut-residing GBS to cause sepsis in a murine model of LOS is still unknown. Here we infected five-day-old mice by oral gavage with different strains of GBS, but we did not observe GBS translocating the intestinal epithelium in the pups, unlike E. coli. We next induced EOS in newborn mice by vaginally inoculating pregnant mice to assess GBS dissemination and compare the EOS model to our LOS model. After in utero infection, most pups were intestinally colonized with GBS and, interestingly, only pups with GBS colonies in the lungs had systemic GBS infections. This data suggests neonates are at risk of GBS infection in utero, but not following delivery, even if GBS colonizes the neonatal intestine. This work could elucidate the pathogenesis of EOS as compared to LOS, leading to a deeper understanding of modes of GBS transmission.

Supported by NIH: DK122187
Antigen-specific nanoparticle tolerance treatment actively induces both FoxP3- and IL-10-dependent regulatory mechanisms
Authors: Podojil Joseph R , Cogswell Andrew , Chiang Ming-Yi , Neef Tobias , Murthy Tushar , Boyne Michael , Elhofy Adam , Miller Stephen D
Published: January 2023

There are many autoimmune diseases where pathologic self T cells are the underlying cause of disease symptoms and progression. T cells are found at the site of tissue destruction, resulting not only in the exacerbation of disease, but also the release of self-epitopes in the context of inflammation resulting in the activation of additional T cell populations perpetuating disease. Published data have shown treatment with antigen-containing biodegradablepoly(lactide-co-glycolide) (PLGA) nanoparticles, i.e. tolerogenic immune-modifying particles (TIMP), is both safe and induces antigen-specific tolerance in mouse models of autoimmunity and allergy, as well as in a celiac disease Phase I/IIa clinical trial. This study further investigated the role and mechanism of TIMP-induced tolerance. The present data show that TIMP treatment modulated T cells specific for spread epitopes associated with disease progression, but not encapsulated within the TIMP, i.e. tissue-specific bystander suppression. The PLP139–151 TCR transgenic model systems was utilized to determine the cellular and molecular mechanisms driving antigen specific tolerance mediated by tolerogenic nanoparticle treatment. These data show antigen-specific TIMP treatment induced both FoxP3+ iTregs and IL-10+ Tr1 regulatory phenotypes within the antigen-specific T cell populations in both naïve mice and mice pre-primed with antigen/CFA. Additionally, both functional Tregs and IL-10 are required for TIMP-induced tolerance. Treatment thus activates various antigen-specific Treg subsets capable of regulating responses to disease-relevant autoepitopes not encapsulated within the TIMP via release of immunoregulatory cytokines within the inflammatory site.

Supported by funding provided by Cour Pharmaceuticals Development, Inc.
Liver macrophage subsets differentially regulate metastasis in pancreatic cancer
Authors: Thomas Stacy K , Beatty Gregory L
Published: January 2023

Metastasis is the main cause of morbidity and mortality in patients with cancer. In pancreatic ductal adenocarcinoma (PDA), the liver is the most common site of metastasis. During PDA progression, the liver microenvironment becomes altered to support metastatic seeding and colonization by disseminated tumor cells (DTC). Macrophages (MΦ), including liver-resident Clec4f+ Kupffer cells (KCs) and bone marrow derived MΦ (BMDMs), are the dominant immunological components of this niche, yet their precise roles in regulating metastasis remain poorly defined. Here, we sought to understand the interaction of DTCs with liver MΦ as they seed and colonize the liver. To model liver metastasis, mice received intraportal (iPo) injection of murine PDA cells with analysis 2 or 14 days later to examine metastatic seeding and outgrowth, respectively. Liver metastatic burden and immune cell populations were quantified by flow cytometry and immunohistochemistry. Liver MΦ were depleted using clodronate encapsulated liposomes (CEL) and KCs were depleted using Clec4fDTR mice administered diphtheria toxin. DTCs were found to interact with BMDMs more often than KCs during the seeding phase of metastasis. During colonization, KCs became excluded from metastatic colonies whereas BMDMs actively infiltrated. Although depletion of liver MΦ and KCs specifically did not affect metastatic seeding, MΦ depletion during the colonization phase inhibited metastatic colony outgrowth. Interestingly, BMDM infiltration into lesions was largely unaffected by CEL treatment, whereas KCs were uniformly depleted. This finding suggests that MΦ residing outside of metastatic lesions may be critical to supporting metastatic outgrowth.

Supported by grants from NIH (T32 HL0439, F30 CA257287, R01 CA245323)
The role of complement-mediated signaling during antigen presentation
Authors: Bird Caryn Lishele , Pacholczyk Gabriela , Johnson Theodore
Published: January 2023

Local production of complement C3a and C5a at the immune synapse by activated dendritic cells (DCs) enhances T cell responses in a variety of experimental systems. We have shown that complement C3 plays a mechanistic role in driving synergistic anti-tumor effects of blocking indoleamine 2,3-dioxygenase (IDO) during radiation and chemotherapy in a murine brain tumor model. We hypothesize that, when anti-tumor responses are allowed to happen, locally produced complement C3 acts as a costimulation factor for activation of T cells. To study the role of complement C3 in promoting T cell responses against a nominal tumor antigen (gp10025–33 peptide expressed by B16F10 melanoma), we used T cells from syngeneic pmel-1 mice (TCR transgenic with CD8 T cells that recognize the cross-presented gp10025–33 peptide). We found florid deposition of complement C3 in B16F10 tumors when IDO-blockade was added to a pmel-1/gp10025–33 based vaccine. To model homeostatic conditions of the tumor microenvironment, we used an in vitro co-culture system with gp10025–33 peptide in stringent limited supply. Under these conditions, pmel-1 CD8 T cells stimulated by DCs from complement C3 knock-out (C3-KO) mice had decreased activation marker expression, proliferation, and cytokine effector function, relative to pmel-1 CD8 T cells stimulated by DCs from wild-type (WT) mice. These findings suggest that suppression of local complement production by DCs in the tumor microenvironment may inhibit anti-tumor immune responses.

Supported by grants from Alex’s Lemonade Stand Foundation, Press On Foundation, and Hyundai Hope On Wheels.
Impact of constitutive FLT3 signaling on dendritic cell development and function in a genetically-engineered mouse model of Acute Myeloid Leukemia
Authors: Flynn Patrick A , Coy Jesse L , Romine Kyle Alexander , Kosaka Yoko , Lind Evan
Published: January 2023

Acute myeloid leukemia (AML) is a hematological cancer with a very poor prognosis. FLT3-ITD mutations that cause constitutive FLT3 signaling are commonly seen in patients. Our lab and others have shown that AML patient samples exhibit hallmarks of immune exhaustion such as T cell dysfunction, increased MDSCs, and increased Tregs that associate with worse survival. In solid tumor models DCs have been shown to be important for anti-tumor T cell activity and we aim to further describe the role of DCs in the immune response to leukemia. FLT3 signaling is critical for DC development but the effects of FLT3-ITD on DCs and how it contributes to leukemogenesis remains unclear. In a novel FLT3-ITD driven AML mouse model we found that DCs from these mice have elevated pFLT3, indicating constitutive FLT3 signaling. Moreover, significantly expanded DC progenitors and DCs in the bone marrow and spleen are observed. Analysis of serum cytokines identified increased levels of IL-27, IL-10, and IL-17a in leukemia mice compared to healthy controls, suggesting that the expansion of DCs in these leukemia mice may be driving pathogenic expansion of T helper subsets. Furthermore, we measured higher levels of circulating Th2, Th17, and Treg phenotypes in the blood compared to healthy controls. Thus, we hypothesize that FLT3-ITD DCs are a significant contributor to T cell skewing in AML resulting in poor anti-tumor responses. We are using scRNA-seq methods to identify transcriptional changes in FLT3-ITD DCs that lead to the phenotypes we identified. Understanding how FLT3-ITD DCs contribute to AML immune suppression is critical to interpreting leukemia etiology and the development of targeted therapies such as immune checkpoint blockade or small molecule inhibitors.
Long-term sterile immunity induced by an adjuvant-containing live-attenuated AIDS virus
Authors: Yasutomi Yasuhiro
Published: January 2023

Antigen 85B (Ag85B) is one of the most dominant proteins secreted from most mycobacterial species, and it induces Th1-type immune responses as an adjuvant. In the present study, a new strategy to improve live-attenuated SHIV was examined by genetically engineered virus that co-expresses an immunostimulatory molecule Ag85B such as an adjuvant. We genetically constructed a live nef-deleted simian-human immunodeficiency virus to express the adjuvant molecule Ag85B (SHIV-Ag85B) and assessed vaccine effects in cynomolgus macaques. The SHIV-Ag85B could not be detected 4 weeks after injection in cynomolgus macaques, and strong SHIV-specific T cell responses were induced in these macaques. Moreover, most of macaques inoculated with SHIV-Ag85B showed protective effects against the intravenous challenge of pathogenic SHIV89.6P, and viral RNAs remained undetectable for more than 200 weeks. Remarkably, eradication of SHIV89.6P was confirmed by an adoptive transfer experiment and CD8+ cell depletion study. In these macaques, SHIV antigen-specific CD8+ T cell responses with polyfunctionality were rapidly induced in the acute phase of SHIV89.6P challenge. The results suggest that SHIV-Ag85B elicited strong sterile immune responses against pathogenic SHIV and that it may lead to the development of a live vaccine and cure for AIDS virus infection.

Supported by Health Science Research Grants from the Ministry of Health, Labor and Welfare of Japan, Ministry of Education, Culture, Sports, Science and Technology, Japan, Japan Agency for Medical Research and Development, AMED (19ak0101047h0004, 19fk0410025h0001, 19ak0101068h0003 and 19fk0410011h0702).
Exposure to SARS-CoV-2 spike protein induces antigen specific IgE and basophil activation
Authors: Zhang Tianyi , Magazine Nicholas , McGee Michael Conrad , Huang Weishan
Published: January 2023

The spike glycoprotein of SARS-CoV-2 (S) has been used as a main target antigen for vaccine design. There are concerns of vaccine-induced allergies, but mostly about allergic responses to the adjuvant. We, however, found that repetitive exposure to SARS-CoV-2 spike, not Nucleocapsid (N) protein, can induce robust type 2 inflammation in the airway of mice, which is exacerbated by the presence of house dust mite allergens. This type 2 inflammation is accompanied with elevated IgE and neutralizing antibody production. To further test whether spike glycosylation influences these responses, we compared the immunogenicity of insect cell expressed spike (in-S), mammalian cell expressed spike (m-S) and deglycosylated spike (dg-S). With intranasal treatment, in-S induced high levels of anti-spike/S1 specific IgE, while m-S and dg-S induced detectable NAb titers as determined by pseudo-virus (PV) neutralization assays. Moreover, exposure to in-S induced a significant amount of IgE+ basophils in the spleens and lungs of the mice. These spike-induced phenotypes are dependent on the TLR4/MyD88 and STAT6 signaling pathways. When delivered via intramuscular immunization following the priming/boosting protocol, spike protein induced high levels of antigen-specific IgE and IgE+ basophils. Of note, in-S induced the highest level of anti-spike/S2 IgE. However, interestingly, deglycosylation of spike antigen resulted in enhanced induction of NAbs and spike-specific CD8+ T cells following intramuscular immunization. Together, our data suggests that vaccination with SARS-CoV-2 spike may induce antigen-specific allergic response, while immunization with the deglycosylated spike protein may help enhance protective immune responses.

Supported by National Institutes of Health (R01 AI151139, R56 AI146226 and P20 GM130555). W. Huang also received research support from MegaRobo Technologies Corporation.
Hybrid immunity and vaccine breakthrough lead to robust humoral response and antibodies that effectively neutralize SARS-CoV-2 variants
Authors: Bates Timothy A , McBride Savannah K , Leier Hans C , Lyski Zoe L , Messer William B , Curlin Marcel E , Tafesse Fikadu G
Published: January 2023

Current COVID-19 vaccines significantly reduce overall morbidity and mortality and are vitally important to halting the pandemic; individuals who previously recovered from COVID-19 display enhanced immune responses after vaccination (hybrid immunity) compared to their naïve-vaccinated peers. However, the effects of vaccine breakthrough infections on humoral immune response remain to be determined. Here, we measure neutralizing antibody responses from 104 vaccinated individuals including those with breakthrough infections, hybrid immunity, and no infection history. We find that human immune sera collected after either a breakthrough infection or a vaccination post-natural infection will both broadly neutralize SARS-CoV-2 variants to a similar degree. While age negatively correlates with antibody response after vaccination alone, no such association was found in breakthrough or hybrid immune groups. Together, our data suggest that the additional antigen exposure from natural infection substantially boosts the quantity, quality, and breadth of humoral immune response regardless of whether it occurs before or after vaccination.

Supported by grants from the M.J. Murdock Charitable Trust, OHSU Innovative IDEA grant (1018784), NIH (R01AI145835, T32HL083808), and an unrestricted grant from the OHSU Foundation.
Characterizing oncolytic potential and safety of a G-protein pseudotyped vesiculovirus in sarcoma
Authors: Dumbauld Chelsae R , Seetharam Mahesh , Nagalo Bolni Marius , Borad Mitesh
Published: January 2023

Several vesicular stomatitis virus (VSV)-based oncolytic therapies have advanced to phase II clinical trials for treatment of solid tumors. As most antibodies produced after VSV infection are directed towards the surface glycoprotein (G), switching this G-protein of VSV to that of a novel virus could enable repeat dosing in cancer patients by evading adaptive immunity. We aim to study whether a hybrid virus expressing the G-protein of a closely related and non-pathogenic vesiculovirus exhibited oncolytic efficacy across sarcoma models.

We compared cytotoxic effect of VSV, VX1 (wild-type novel virus), and VxG (hybrid) post infection in sarcoma lines. We quantified viral progeny production over time and compared real-time rate of viral-induced apoptosis. Sarcoma lines were tested for production of type-I interferon (IFN) and protection from VxG-induced cell death by exogenous IFNβ. We completed in vivo efficacy studies of intratumoral administration of VxG in xenograft and syngeneic models. From syngeneic tumors, we performed immunophenotyping on residual tumors treated with VxG compared to controls.

VxG was observed to be well tolerated and efficacious. In a xenograft model, a single low-dose administration of VxG completely inhibited tumor growth compared to controls. Lower tumor burden was observed in syngeneic tumors treated with VxG compared to controls. Comparisons of tumor infiltrating lymphocytes between residual VxG treated and control tumors will be completed.

We conclude that the novel hybrid virus VxG has potent oncolytic effects in multiple models of sarcoma and low toxicity in immunocompetent mice. Future endeavors will focus on improved viral engineering and further pre-clinical studies.

Supported by the Mayo Clinic Graduate School of Biomedical Sciences
Novel RNA-based Cancer Vaccine for AML immunotherapy
Authors: Liu Haiyan , Tham Guo Hong , Ren Xi , Yang Henry
Published: January 2023

Acute Myeloid Leukemia is a rapidly progressing malignancy relying primarily on chemotherapy as the frontline treatment option. Success of personalized neoantigen vaccines in melanoma patients has garnered great attention in the field of immunotherapy. We hypothesize that personalized cancer vaccine, in the form of mRNA, can be a viable treatment strategy for AML patients. Next generation sequencing was performed on both C57BL6Jinv mouse’s and C1498 cell line’s nucleic acid materials. Three different neoepitope prediction pipelines were utilized to identify potential immunogenic neoepitopes. Candidate neoepitopes within intron polyadenlyation sites (iPAS), retained intron (RI) sites and somatic mutation (SM) sites were found. A total of 285 SM-derived neoepitopes were identified. Top 38 highly expressed genes were selected for further analysis. Using Sanger sequencing, we confirmed 37 out of the 38 mutations. A total of 5 iPAS-derived neoepitopes were identified. A total of 26 RI-derived neoepitopes were identified in our first RNAseq analysis and a separate 476 RI-derived neoepitopes were identified in our second RNAseq analysis. With these three categories of candidates, we narrowed down our selection and did a preliminary screen for immunogenic epitopes using a total of 71 synthetic peptides in an ELISpot assay. Conventional ELISpot assay’s results were however non-reproducible, thereby propelling us to adopt Culture ELISpot assay. Subsequently, 10 neoepitope candidates were short-listed for further in-vivo evaluation. mRNA vaccine was synthesized and evaluated in a murine AML model. T cell responses against the 10 neoepitopes were analyzed to provide pre-clinical evidence for patient studies.

Supported by NMRC CS-IRG grant
Immunogenicity and efficacy of a multiple antigen-presenting system (MAPS™) COVID-19 vaccine in Non-human Primates
Authors: Besin Gilles , Cieslewicz Brian , Makrinos Daniel , Haridas Renuka , Burke Heidi , Stevenson Taylor , Malley Richard
Published: January 2023

In response to the COVID-19 pandemic, the international scientific and biopharmaceutical community has rallied with unprecedented speed to develop effective vaccines to combat this novel pathogen. Despite this remarkable success, the rise of SARS-CoV-2 variants, some of which appear more resistant to the effects of vaccination, highlights the potential need for additional COVID vaccines.

We used the MAPS technology, in which proteins are presented on a polysaccharide polymer to induce antibody, Th1, Th17 and CD8+ T cell responses, to engineer a COVID-19 vaccine. The COVID-19 MAPS vaccine contained a fragment of the S-RBD sequence of the original strain. Nonhuman primates (NHP) were immunized twice with the COVID-19 MAPS for assessment of immunological responses. NHPs were then challenged with SARS-CoV-2 and protection was assessed by measuring the viral load in the lung and in the nasopharynx.

The COVID-19 MAPS vaccine generated robust neutralizing antibodies in all three animal species as well as Th1, Th17 and cytotoxic CD8 T-cell responses in NHPs. Following viral challenge, MAPS-immunized NHPs had significantly lower viral load both in the nasopharynx and the lung than control animals. This difference was appreciable in the earliest stages following challenge.

A COVID-19 MAPS vaccine containing the RBD of the D614G SARS-CoV-2 strain was very immunogenic in mice, rabbits and NHPs, generated de novo T cell responses and was highly protective against viral challenge in NHPs. These results, as well as the plug-and-play nature of the MAPS technology, in which different RBD variants can be included in a single vaccine preparation, suggest that the MAPS platform could be used to make a vaccine to target multiple variants of SARS-CoV-2.
The Reaction of Normal and Rabbit Pox Immune Adult Rabbits to Vaccination with Vaccine Virus
Authors: Rosahn Paul D. , Hu C. K. , Pearce Louise
Published: January 2023

Summary and Conclusions

Vaccination with vaccine virus of a rabbit colony of approximately 1500 animals was carried out as a prophylactic measure against rabbit pox. The observations made on the adult population are discussed in the present paper.

Of 233 presumably susceptible animals, 195 or 83.7 per cent gave a positive local reaction to vaccination. Generalized lesions, comprising in particular cutaneous and mucocutaneous papules, adenopathy, and in males an orchitis, developed in 78 animals, an incidence of 33.5 per cent. Nine of these cases had had a negative vaccination. Clinical evidence of susceptibility to vaccinal infection was thus demonstrated in 204 presumably susceptible rabbits, an incidence of 87.6 per cent.

Of 310 rabbits which had recovered from rabbit pox ten months previously and were, therefore, presumably refractory to vaccination with vaccine virus, 294 or 92.3 per cent gave completely negative results. A positive reaction to vaccination developed in 21 animals, however, and there were 3 cases of generalized lesions in animals with a negative local vaccination site, a result which shows that the refractory state induced by rabbit pox may eventually disappear.

An outstanding feature of the results concerned the different character of the reaction of female as compared with male rabbits. A slight, even a negative, local reaction to vaccination was often observed in females and there were comparatively few cases of generalized lesions. This type of reaction was especially striking in the case of pregnant animals. The differences between the lower incidence of both local and generalized lesions in pregnant rabbits as compared with males were found to be statistically significant.
Some Factors Which Affect the Ultrafiltration of Antipneumococcal Sera
Authors: Goodner Kenneth , Horsfall Frank L. , Bauer Johannes H.
Published: January 2023

Biological and physical evidence has been advanced to support the view that aggregation and dispersion are involved in the behavior of antibodies.
Studies of the Early Stage of Resistance to Pneumococcic Infection
Authors: Schlesinger Helen , McBroom Josephine
Published: January 2023

The early stage of resistance to infection by a type I pneumococcus may be elicited within 48 to 72 hours in response to intracutaneous treatment with the homologous type I vaccine, but not in response to the heterologous type III vaccine.

Small quantities of mouse-protective substances are demonstrable in the blood of slightly more than half of the resistant animals by the sixth and tenth days following a single intracutaneous immunizing injection; they are not present in demonstrable quantities as early as the second and third days following this treatment. No increase in agglutinins could be demonstrated during the early stage of resistance.
Sensitization to Ragweed-Extract and the Production of Antibodies by Means of Adjuvants
Authors: Kulka Anna M. , Hirsch Dorothy
Published: January 2023

Summary and Conclusions

Ragweed-pollen extract combined with a lanolin-like substance and paraffin oil containing killed tubercle bacilli induces sensitization and antibody-formation against pollen-extract far more vigorously than pollen-extract alone. Sensitzation in both groups of animals was noted earlier than the presence of circulating antibodies. Animals with the highest cutaneous sensitiveness had also the highest precipitative titer. The presence of passively transferable antibodies against ragweed-pollen could be demonstrated in animals injected with pollen-extract plus adjuvants.
Weather and Susceptibility in Relation to the Spread of Common Cold; Effect of Ascorbic Acid, in Massive Dosage, on Duration
Authors: Brown WM. B. , Mahoney F. , Niedringhaus A. , Locke Arthur
Published: January 2023

A study was made of the spread of common cold through a group of 1600 girls of college age, as affected by the weather, use of cigarettes, and relative susceptibility to common cold; of the season-to-season changes in physiological fitness as appraised by a modified Flack test; and of the effect, on duration of common cold, of early recognition and care, and ascorbic acid in massive dosage.

Sharp increases in number of new colds per day occurred during periods of falling temperature accompanied by heavy rain. The increases began and were most steadily sustained in the girls with a total of 4 or more colds for the year. The smokers, also, had a tendency to precede the nonsmokers. The weather-precipitated rises in colds-incidence tended to begin 24 to 48 hours after the beginning of the weather change and to persist for 24 to 48 hours after its cessation. The total number of colds, for the September wave, reached its halfway-point 2 days earlier for the more-susceptible fraction than for the less-susceptible fraction.

Only a slight deterioration in average physiological fitness occurred with the progress of the year. The deterioration was not uniform but restricted to a small fraction, partially balanced by a smaller group showing improvement. The smokers appeared to be slightly less fit, on the whole, than the nonsmokers. Nose involvement predominated in the fall colds and throat involvement in the winter colds which were, mainly, seconds and thirds rather than first colds.

A total of 298 colds was studied for the ascorbic acid appraisal; 119 of which were controls given placebos of citric acid. Approximately 60 per cent of the colds with throat involvement only appeared to respond to the basic management given both the control and ascorbic acid groups—restriction of activity, precaution against chilling, and increased fluid intake—with a spontaneous checking of further development. The colds with nose involvement were equally responsive to management, alone, if recognized and submitted to care within 7 hours of presumed onset. For those not recognized and submitted to care until 8 to 28 hours after presumed onset, the percentage of spontaneous checking of further development was, for those given one gram of ascorbic acid at time of first examination and again 24 hours later, 59 ± 5 and, for those given citric acid, 21 ± 4.
Combined Active—Passive Re-Immunization against Tetanus in Previously Immunized Individuals: Experimental and Clinical Evidence
Authors: Miller John J. , Ryan Mary Louise
Published: January 2023

Combined active-passive re-immunization against tetanus was studied in 29 previously actively immunized rabbits by means of tetanus antitoxin titrations. Both fluid and alum-precipitated tetanus toxoids in doses of 0.05 or 0.1 ml were used. Commerical tetanus antitoxin was employed in a dose of 150 units (except as in (c) below). When these doses of antitoxin and toxoid were injected into opposite extremities, the initial increase in titer due to the injected antitoxin was followed in 3 to 6 days by a higher and prolonged secondary increase. This latter could only be a true secondary immune response. When toxoid and antitoxin were mixed in the same syringe no response occurred. Increasing the dose of separately injected antitoxin 3.3 fold led to higher initial titers but did not suppress the secondary immune response. The addition of hyaluronidase to the toxoid did not speed the secondary immune response but appeared to increase the total amount of homologous antitoxin produced by the animals. Four previously immunized individuals, two children and two adults, were studied following accidental wounding. They were injected with the usual prophylactic dose (1500 units) of tetanus antitoxin and, in the opposite extremity, either fluid or alum-precipitated tetanus toxoid. Following initial increases in titer due to the antitoxin, three of the four exhibited secondary immune responses to the toxoid 4 to 5 days later.
Q Fever in California
Authors: DeLay Paul D. , Lennette Edwin H. , DeOme Kenneth B.
Published: January 2023

Coxiella burneti, the causal rickettsia of Q fever, has been recovered from dustladen air on a dairy in Southern California, and from dust-laden air on a sheep ranch in Northern California.

The signficance of these recoveries of the organism from a contaminated environment is discussed.
The Effect of Human Serum Upon the Agglutination of Enzyme-Treated Erythrocytes
Authors: Hoyt Robert E. , Zwicker Helen
Published: January 2023

Since Morton and Pickles (1) showed that enzyme-treated red cells are agglutinated by “incomplete” antibodies in the absence of hydrophilic colloids the observation has been put to practical use in the detection of “incomplete” antibodies in the maternal blood. In comparing techniques for demonstrating “incomplete” antibodies, using pooled human serum, 30% bovine albumin and enzyme-treated group O, Rh positive erythrocytes, Kuhns and Bailey (2) found the method using papain-treated cells to be the most sensitive, with pooled human serum as a diluent giving the lowest titres. Other reports are in general agreement and it is believed that the test using enzyme-treated cells gives reliable information regarding the presence of “incomplete” anti-Rh antibodies.

In a study of the agglutination of enzyme-treated red cells we have compared the titres of anti-Rho agglutinating sera and anti-Rho blocking sera when measured against treated and untreated cells.
The Effect of Trypsin on Hemagglutination by Murine Encephalomyelitis Virus (GDVII)
Authors: Morris Marion C.
Published: January 2023

The GDVII strain of murine encephalomyelitis virus has recently been shown to cause agglutination of human red cells at 4 C (1). This reaction did not occur at room temperature or at 37 C, and the agglutination occurring at 4 C was dispersed when the temperature was raised. This reversal of hemagglutination was due to elution of the virus but was not accompanied by a destruction of cell receptors as occurs with viruses of the mumps-influenza group. Of a variety of red blood cells examined, only human cells were found susceptible to agglutination by GDVII virus.

The report of Rheims, Dodd and Hudson (2), that treatment of human group O cells with trypsin increased their agglutinability by mouse lung infected with PR8 influenza virus, suggested the present study. In anticipation of the results to be reported, it may be said that exposure of the cells to trypsin did not affect hemagglutination by GDVII virus.
Studies on Bacterium Tularense Antigens
Authors: Ormsbee Richard A. , Larson Carl L.
Published: January 2023

An investigation of the chemical and physical characteristics of purified B. tularense antigen preparations indicated the presence of at least four and possibly six antigenic components probably derived from the cell wall. The treatment of aqueous suspensions of whole organisms with ethyl ether, and the purification of the extract by salt precipitation, differential centrifugation, and dialysis produced a product which possessed uniform immunogenic activity and reproducible ratios of the various chemical constituents. Electron microscope photographs of such a purified preparation indicated that the material was indistinguishable in electron density from pure cell-wall material prepared by other methods. Lipid extraction of purified preparations yielded an immunogenically active preparation composed of a polysaccharide and an amino acid complex and containing organic P. The reducing sugar value found was almost identical with the calculated value for N-acetyl-glucosamine. Various qualitative tests suggest that the N-acetyl derivative of D-glucose (-D-glucopyranoside) may be the sugar resulting from acid hydrolysis of the polysaccharide moiety. The lipid-free purified antigen preparations were indistinguishable from the initial crude extracts in their immunogenic or serologic activity, except for the greatly increased activity per unit weight of antigen.
Immunological Properties of a from of Non-Precipitating Diphtheria Antitoxin Which Does Not Sensitize Human Skin
Authors: Kuhns William J.
Published: January 2023

A type of human non-precipitating diphtheria antitoxin is described which is not able to cause immediate wheal reactions in appropriate recipients. Individuals who form this antitoxin following a booster dose of toxoid develop no immediate reactivity upon subsequent testing with intradermal toxoid.

The sera of ten persons who formed only non-precipitating non-skin-sensitizing antitoxin were studied in detail and the following characteristics were observed. No fixation of complement occurred unless large amounts of antitoxin were used, and specific coprecipitation of antitoxin in two high titered sera resulted upon the addition of precipitating antitoxin and toxin. None of the sera tested were able to sensitize human skin, nor was there any evidence that this antitoxin remained at intradermal sites. When three sera containing the highest titers were separated electrophoretically, the antitoxin was found to migrate as a slow-moving gamma globulin (γ2). Three of seven sera were able to inhibit immediate reactions at sensitized sites when combined with equivalent amounts of toxoid. Four sera could not be evaluated for this property because they had an irritative effect when injected alone in the skin.
Antigenicity of β-Propiolactone-Inactivated Virus Vaccines
Authors: LoGrippo Gerald A. , Hartman Frank W.
Published: January 2023

A simple and rapid procedure for the preparation of inactivated virus vaccine is described using β-propiolactone (BPL), an agent not previously employed in the preparation of vaccines. β-propiolactone is capable of inactivating viruses in 10–15 minutes at 37°C whereas formalin and phenol require days for the same degree of inactivation under parallel conditions. Satisfactory inactivated vaccines can be prepared with eastern equine encephalomyelitis virus, rabies virus and MM-virus with the use of BPL. A significantly higher degree of antigenicity is demonstrated with β-propiolactone-inactivated virus vaccines than with formalin- and phenol-inactivated virus vaccines. Two to four times the minimal effective viricidal concentration of BPL can be used in the preparation of virus vaccines without critical loss in antigenicity.
An Investigation of Some of the Factors Determining the Decrease in Complement Activity in Anaphylactic Shock
Authors: Rice Christine E.
Published: January 2023

Guinea pigs actively sensitized to egg albumin mixed with adjuvants developed more acute symptoms of shock and showed a more pronounced and rapid fall in complement titer when challenged intravenously 6 weeks later than did similar groups sensitized to egg albumin without adjuvants. The change in C′ titer tended to be greatest in the guinea pigs exhibiting very severe respiratory symptoms.

Fatal protracted shock followed the intraperitoneal injection of large amounts of antigen in highly sensitized guinea pigs, but the decrease in complement activity was usually less pronounced than in the intracardially-challenged animals dying within a 10-minute interval.

Actively and passively sensitized guinea pigs in which fatty livers and a marked fall in C′ titer had been induced previously through repeated injection of ethionine, exhibited respiratory symptoms as acute as the sensitized controls. Less severe respiratory symptoms were recorded on challenge in actively-sensitized guinea pigs that had been treated with gum acacia for several weeks to induce low C′ titers; symptoms of initial excitement with subsequent weakness were more conspicuous. In both groups of sensitized guinea pigs, complement titers declined further on challenge.

In passively sensitized guinea pigs, challenged soon after intraperitoneal injection of antiserum, the change in C′ titer was negligible, even though some of these animals developed symptoms of slight to moderate shock. A rapid and striking fall in C′ titer occurred in guinea pigs of the same groups that experienced acute shock following intravenous challenge 18 hours to 7 days after injection of antiserum.
The Effect of Ficin on the Agglutination of Human Red Blood Cells
Authors: Makinodan Takashi , Macris Nicholas T.
Published: January 2023

A concentration of 1.5 mg of ficin per ml of packed cells, a pH range from 4.7 to 7.8, an incubation temperature of 22° to 37°C and an incubation period of 10 to 30 minutes were found to be the optimum conditions for making A, B and Rh-positive cells agglutinable by the homologous “complete” and “incomplete” antibodies. Ficin was found to be more efficient than trypsin and papain for increasing the sensitivity of O Rh-positive, A and B cells in hemagglutination. The intracellular protease activator, L-cysteine-HCl, accelerated significantly the rate of sensitization of O Rh-positive cells by papain, but not by ficin.
Agglutinin Production in Normal, Sublethally Irradiated, and Lethally Irradiated Mice Treated with Mouse Bone Marrow
Authors: Makinodan Takashi , Gengozian Nazareth , Congdon Charles C.
Published: January 2023

Maximum agglutinin production was attained when the mice were 12 weeks of age. The optimum antigen dose to induce the maximum agglutinin response was found to be 1 ml of a 1.0% RBC suspension. Isologous bone marrow suspension protected 95% of the lethally x-irradiated mice (950 r). The ability to produce sheep agglutinin was recovered 30 days after the lethal irradiation-bone marrow treatment, whereas 60 days were required to produce rat agglutinin.
The Detection of Antibodies in Hayfever Sera by Means of Hemagglutination
Authors: Feinberg Richard J. , Davison Joan D. , Flick John A.
Published: January 2023

Antibodies in the sera of hayfever patients against pollen antigens can be measured, in vitro, by adsorbing the antigen onto tanned erythrocytes and using such antigen-coated cells with the patient's serum in a hemagglutination test. This technique detects antibodies mainly in the sera of hayfever patients who have received specific hyposensitization, but occasional untreated hayfever subjects also possess antibody detectable by this method. Probably both multivalent- and univalent-acting types of antibodies can be measured by this technique. Evidence is presented which indicates that the hemagglutinating antibody found in these hayfever sera is not identical with the reagin type of antibody.
Integration of Human Serum and Serum Fraction Diffusion Patterns
Authors: Glenn William G. , Garner Anna C.
Published: January 2023

Direct photometry of human serum and plasma fractions, using the serum agar measuring integrator (SAMI), is described.

In diffusion patterns of human serum reacting with anti-human rabbit sera, the fastest zone was consistently an albumin system and the next major dense zone was β globulin.

A discussion of the zonal characteristics of albumin and γ globulin is included.
The Elimination of Heterologous Serum Proteins and Associated Antibody Responses in Guinea Pigs and Rats
Authors: Weigle William O. , Dixon Frank J.
Published: January 2023

The rate of elimination of I* protein antigens was used as a method of observing immune responses in guinea pigs and rats when single, spaced injections of antigens (7.5 mg/kg) were administered. Animals exhibiting immune responses eliminated the labelled antigens at accelerated, immune rates similar to those described in rabbits. Also, the relationship of the rapid, immune elimination of antigen to the subsequent appearance of antibody was similar to that seen in the rabbit. However, the immune responses to soluble protein antigens in guinea pigs had longer induction periods and slower immune elimination of antigen than in rabbits.

The slower immune elimination of antigen in guinea pigs than in rabbits was presumably the result of a slower appearance of antibody in guinea pigs since antigen-antibody complexes once formed were rapidly eliminated. In guinea pigs injected with I*BSA and I*RSA, circulating antigen-antibody complexes could only be demonstrated during the phase of rapid, immune antigen elimination.

On the basis of observations in the guinea pig and rat, there is no apparent correlation between the type of protein antigen and the rate of nonimmune elimination or the length of the induction period prior to antibody synthesis.

Determination of the rates of elimination of antigen was found to be as sensitive a method of testing for an immune response to BGG, HGG, and RGG in guinea pigs as was anaphylaxis.
Studies on the Viral Spectra of Tissue Culture Lines of Human Cells
Authors: Deinhardt Friedrich , Henle Gertrude
Published: January 2023

A comparison was made of 12 tissue culture cell lines of human origin with respect to their susceptibilities to a limited number of viruses. On the basis of the viral spectra obtained the lines could be divided into 2 groups corresponding to those established in morphological grounds. The observed interactions of the various viruses with the different cells fell into 4 categories: (1) viral multiplication with development of cytopathogenic effects; (2) propagation of virus in the absence of cellular destruction; (3) cytopathogenicity without viral replication; and (4) no growth of virus nor cellular damage. The implications of some of these findings have been discussed.

Only cells of one of the groups (I) were susceptible to poliomyelitis, Coxsackie B and adenoviruses. None within group I were more sensitive to poliomyelitis than HeLa cells.

Inoculation of cells of both groups with specimens from cases of infectious or serum hepatitis failed to yield evidence of infection.