Lungs Of Syrian Golden Hamsters Research Articles

Antiviral Immunoglobulins of Chicken Egg Yolk for Potential Prevention of SARS-CoV-2 Infection.

Background: Some viruses cause outbreaks, which require immediate attention. Neutralizing antibodies could be developed for viral outbreak management. However, the development of monoclonal antibodies is often long, laborious, and unprofitable. Here, we report the development of chicken polyclonal neutralizing antibodies against SARS-CoV-2 infection. Methods: Layers were immunized twice with 14-day intervals using the purified receptor-binding domain (RBD) of the S protein of SARS-CoV-2/Wuhan or SARS-CoV-2/Omicron. Eggs were harvested 14 days after the second immunization. Polyclonal IgY antibodies were extracted. Binding of anti-RBD IgYs was analyzed by immunoblot and indirect ELISA. Furthermore, the neutralization capacity of anti-RBD IgYs was measured in Vero-E6 cells infected with SARS-CoV-2-mCherry/Wuhan and SARS-CoV-2/Omicron using fluorescence and/or cell viability assays. In addition, the effect of IgYs on the expression of SARS-CoV-2 and host cytokine genes in the lungs of Syrian Golden hamsters was examined using qRT-PCR. Results: Anti-RBD IgYs efficiently bound viral RBDs in situ, neutralized the virus variants in vitro, and lowered viral RNA amplification, with minimal alteration of virus-mediated immune gene expression in vivo. Conclusions: Altogether, our results indicate that chicken polyclonal IgYs can be attractive targets for further pre-clinical and clinical development for the rapid management of outbreaks of emerging and re-emerging viruses.

IMM-BCP-01, a patient-derived anti-SARS-CoV-2 antibody cocktail, is active across variants of concern including Omicron BA.1 and BA.2.

Monoclonal antibodies are an efficacious therapy against SARS-CoV-2. However, rapid viral mutagenesis, led to escape from most of these therapies, outlining the need for an antibody cocktail with a broad neutralizing potency. Using an unbiased interrogation of the memory B cell repertoire of convalescent COVID-19 patients, we identified human antibodies with broad antiviral activity in vitro and efficacy in vivo against all tested SARS-CoV-2 variants of concern, including Delta, Omicron BA.1 and BA.2. Here, we describe an antibody cocktail IMM-BCP-01, that consists of three patient-derived broadly neutralizing antibodies directed at non-overlapping surfaces on the SARS-CoV-2 spike protein. Two antibodies, IMM20184 and IMM20190, directly blocked Spike binding to the ACE2 receptor. Binding of the third antibody, IMM20253, to its cryptic epitope on the outer surface of RBD, altered the conformation of the Spike Trimer, promoting release of Spike monomers. These antibodies decreased Omicron SARS-CoV-2 infection in the lungs of Syrian golden hamsters in vivo, and potently induced antiviral effector response in vitro, including phagocytosis, ADCC, and complement pathway activation. Our pre-clinical data demonstrated that the three antibody cocktail IMM-BCP-01 could be a promising means for preventing or treating infection of SARS-CoV-2 variants of concern, including Omicron BA.1 and BA.2, in susceptible individuals.

Acupuncture for ‘frequent attenders’ with medically unexplained symptoms

<h3>Abstract</h3> Using an unbiased interrogation of the memory B cell repertoire of convalescent COVID-19 patients, we identified human antibodies that demonstrated robust antiviral activity <i>in vitro</i> and efficacy <i>in vivo</i> against all tested SARS-CoV-2 variants. Here, we describe the pre-clinical characterization of an antibody cocktail, IMM-BCP-01, that consists of three unique, patient-derived recombinant neutralizing antibodies directed at non-overlapping surfaces on the SARS-CoV-2 spike protein. Two antibodies, IMM20184 and IMM20190 directly block spike binding to the ACE2 receptor. Binding of the third antibody, IMM20253, to its unique epitope on the outer surface of RBD, alters the conformation of the spike trimer, promoting release of spike monomers. These antibodies decreased SARS-CoV-2 infection in the lungs of Syrian golden hamsters, and efficacy <i>in vivo</i> efficacy was associated with broad antiviral neutralizing activity against multiple SARS-CoV-2 variants and robust antiviral effector function response, including phagocytosis, ADCC, and complement pathway activation. Our pre-clinical data demonstrate that the three antibody cocktail IMM-BCP-01 shows promising potential for preventing or treating SARS-CoV-2 infection in susceptible individuals. <h3>One sentence summary</h3> IMM-BCP-01 cocktail triggers Spike Trimer dissociation, neutralizes all tested variants <i>in vitro</i>, activates a robust effector response and dose-dependently inhibits virus <i>in vivo</i>.

Expression of the Surface Glycoproteins of Human Parainfluenza Virus Type 3 by Bovine Parainfluenza Virus Type 3, a Novel Attenuated Virus Vaccine Vector

Bovine parainfluenza virus type 3 (bPIV3) is being evaluated as an intranasal vaccine for protection against human PIV3 (hPIV3). In young infants, the bPIV3 vaccine appears to be infectious, attenuated, immunogenic, and genetically stable, which are desirable characteristics for an RNA virus vector. To test the potential of the bPIV3 vaccine strain as a vector, an infectious DNA clone of bPIV3 was assembled and recombinant bPIV3 (r-bPIV3) was rescued. r-bPIV3 displayed a temperature-sensitive phenotype for growth in tissue culture at 39 degrees C and was attenuated in the lungs of Syrian golden hamsters. In order to test whether r-bPIV3 could serve as a vector, the fusion and hemagglutinin-neuraminidase genes of bPIV3 were replaced with those of hPIV3. The resulting bovine/human PIV3 was temperature sensitive for growth in Vero cells at 37 degrees C. The replication of bovine/human PIV3 was also restricted in the lungs of hamsters, albeit not as severely as was observed for r-bPIV3. Despite the attenuation phenotypes observed for r-bPIV3 and bovine/human PIV3, both of these viruses protected hamsters completely upon challenge with hPIV3. In summary, bPIV3 was shown to function as a virus vector that may be especially suitable for vaccination of infants and children against PIV3 and other viruses.

Hamster pulmonary endocrine cells with positive immunostaining for calbindin-D28K.

We have examined the distribution of calcium-binding proteins (CaBPs) in adult and fetal lungs of Syrian golden hamsters (Mesocricetus auratus) using immunostaining with confocal laser microscopy and electron microscopy. Single and grouped (neuroepithelial body; NEB) endocrine cells were distributed from bronchi to alveolar ducts in the adult lung. Serial frozen sections immunostained for CaBPs in combination with immunostaining for endocrine markers such as calcitonin gene-related peptide, serotonin, PGP9.5, and synaptophysin revealed that positive immunostaining for calbindin-D28K (CB-D28K) was seen in single endocrine cells and NEBs. However, other so-called EF-hand family CaBPs, parvalbumin and calretinin, were not detected. Electron microscopically, positive immunoreaction for CB-D28K was mainly in the organelle-free cytoplasmic matrix of endocrine cells, and partly in nuclei and associated with secretory granules and endoplasmic reticulum. In fetal developing lungs, endocrine cells appeared first on gestational day 13, and they were positive for all the endocrine markers used. However, pulmonary endocrine cells were positively immunostained for CB-D28K from gestational days 15 and 16 onward. In summary, our observations suggest that CB-D28K is a useful marker for endocrine cells of the lung, and CB-D28K could function as a mediator of endocrine stimulation or calcium homeostasis in pulmonary endocrine cells.

Cystic squamous cell carcinomas in the lungs of Syrian golden hamsters induced by coal oven flue exhaust mixed with pyrolized tar pitch in combination with benzo( a)pyrene

Among a variety of induced pulmonary tumours, cystic squamous cell carcinomas were observed in five Syrian hamsters that inhaled a mixture of pyrolized tar pitch with coal oven flue exhaust (PCE) and additionally received intratracheal injections of benzo(a)pyrene. The histological appearance of these particular tumours is described, compared to similar tumour types in rats and the susceptibility of both species to inert particles is discussed.

Heterogeneity of phagocytosis for inhaled versus instilled material.

Animal experiments involving lung exposure to particles and toxins often use intratracheal instillation, although inhalation is more physiologically relevant to human respiratory tract exposures. Using nontoxic magnetic particles, we examined the particle content distribution within the macrophage population lavaged from the lungs of Syrian golden hamsters exposed either by inhalation or by intratracheal instillation. One to 3 days after iron oxide particle delivery to the lungs, lung macrophages were harvested from animals in pairs, one exposed by inhalation and one exposed by instillation. The macrophages were then magnetically fractionated according to magnetic iron oxide content by flowing the cell suspension through tubing placed within the magnetic field gradient of an electromagnet. Macrophages that had ingested the largest quantity of magnetic iron oxide preferentially collected adjacent to the tubing wall when the electromagnet was at its lowest current; those with fewer particulates collected at correspondingly higher amperages; those cells that were never pulled out of the flowing stream had little or no magnetic iron oxide content. Our results showed that 80% of the lavaged cell population from animals exposed by inhalation had measurable particle content, whereas by instillation, many cells (70%) received no dose at all. Exposure by inhalation produced a more even distribution of dose among lung macrophages. Macrophage intracellular motions showed a decrease when cell content exceeded 7 to 8% of cell volume. We conclude that the method of particle delivery to lungs can influence the dose distribution among lung macrophages.

Immunohistochemical and Biochemical Identification of Pepsinogen Isozymes in the Hamster Lungs: Induction by Polychlorinated Biphenyls

Pepsinogens are acid protease enzymes of pepsin usually found in gastric mucosa. In the present study, we demonstrated the presence of pepsinogen isozymes in male Syrian golden hamster lung tissues by a combined immunohistochemical and biochemical approach. Immunohistochemically, using rat pepsinogen 1 antibody, pepsinogen positive cells were observed mainly in the epithelia of the terminal bronchioles. They demonstrated morphological features of Clara cells. The pepsinogen isozyme pattern of lung tissue determined by polyacrylamide gel electrophoresis was similar to that of stomach mucosa. Treatment of hamsters with polychlorinated biphenyls at a dose of 500 mg/kg body weight ip caused a 2.8-fold increase in pepsinogen content (p less than 0.01) as well as increase in numbers of pepsinogen positive cells in the lung.

Quantification of proliferative lesions in hamster lungs after chronic exposure to cadmium aerosols

The development of proliferative areas in the lungs of Syrian golden hamsters was studied after chronic inhalation of cadmium oxide, cadmium sulfide, cadmium chloride or cadmium sulfate. Lung tissue from randomly selected animals in each group was evaluated by morphometric histopathologic techniques. Estimation of the volumetric ratio of proliferative areas within the lungs of exposed animals showed significantly different extents of these lesions in dependence on the respective cadmium compound administered. The most severe changes were observed after inhalation of cadmium oxide and cadmium sulfide. Lesions were mainly found in the peribronchial region of the lung. Electron microscopic analysis of these proliferative areas revealed that they were composed of ciliated and Clara cells. From its histophatologic appearance this of lesion was qualitatively comparable in all hamsters which had been treated with the different cadmium compounds.

Pulmonary adenoma and endocrine cell hyperplasia in Syrian golden hamster treated with 4-nitroquinoline 1-oxide.

Chronic effects of 4-nitroquinoline 1-oxide (4 NQO) on the lungs of Syrian golden hamsters were studied. 4 NQO was subcutaneously injected weekly for 3 weeks at a dose of 20 mg/kg body weight. The animals were sacrificed at the 65th and 80th experimental weeks. Two cases of pulmonary adenomas were demonstrated in the 10 4 NQO-treated animals at the 80th week, and the tumor cells contained cytoplasmic lamellar inclusion bodies. In a previous study, we reported 4 NQO- induced pulmonary endocrine cell hyperplasias in the 4 NQO-treated hamster after the 20th experimental week (Jpn. J. Cancer Res., 77, 1986). In the present study, 12 pulmonary endocrine cell hyperplasias were recognized in serial sections of the 24 treated animals. The hyperplastic lesions showed positive immunoreactivity to calcitonin. The hyperplastic lesion did not develop to pulmonary endocrine cell neoplasm.

Tumorigenicity of inorganic arsenic compounds following intratracheal instillations to the lungs of hamsters.

The tumorigenicity of arsenic trioxide (As2O3), calcium arsenate (Ca3(AsO4)2) and arsenic trisulfide (As2S3) was studied in male Syrian golden hamsters which received arsenic compounds containing a total dose of 3.75 mg arsenic by means of intratracheal instillations once a week for 15 weeks. As controls, some hamsters were treated with only the vehicle, phosphate buffer solution. During the animals' total life span, one lung adenocarcinoma was seen in the 17 hamsters in the As2O3 group, one lung adenocarcinoma and 6 lung adenomas in the 25 hamsters in the Ca3(AsO4)2 group, one lung adenoma in the 22 hamsters in the As2S3 group and one lung adenosquamous carcinoma in the 21 hamsters in the control group. From these results, we conclude that calcium arsenate is tumorigenic to the lungs of Syrian golden hamsters. The results concerning the other 2 arsenic compounds were not conclusive.

Profiles of steady state levels of messenger RNAs coding for type I procollagen, elastin, and fibronectin in hamster lungs undergoing bleomycin-induced interstitial pulmonary fibrosis.

We have characterized the messenger RNAs (mRNAs) coding for procollagen alpha 1(I), elastin, fibronectin, and actin in the lungs of Syrian golden hamsters by Northern blot analyses. While elastin, fibronectin, and beta-actin were each coded for by a single mRNA species of 4.1 kilobases (kb), 9.1 kb, and 2.1 kb in size, respectively, we identified a major (5.4 kb) and a minor (6.5 kb) procollagen alpha 1(I) mRNA species in the hamster lungs. The mRNAs for the three extracellular matrix proteins showed increased accumulation followed by steady decline in the bleomycin-treated lungs. There were significant differences among the three mRNAs in the relative increase and the time of maximum accumulation. After reaching the peak levels between 2-3 wk posttreatment, the levels of procollagen alpha 1(I) and elastin mRNAs declined to near normal values around the fourth week. In contrast, the accumulation of fibronectin mRNA was maximum in the first week after bleomycin treatment. The procollagen alpha 1(I) mRNA accumulated most dramatically (sevenfold above the levels in the untreated animals) compared with a five-fold increase in mRNA coding for fibronectin. Elastin mRNA increased approximately twofold above the control values. Nuclear runoff transcription experiments demonstrated a selective increase in the rates of transcription of genes coding for procollagen alpha 1(I), fibronectin, and elastin; the extent of transcriptional stimulation of procollagen alpha 1(I) and fibronectin genes was significantly greater than that of elastin. Since the amount of actin mRNA, as well as the rate of transcription of actin gene(s), varied only slightly after bleomycin treatment, we conclude that the metabolism of mRNAs coding for extracellular matrix proteins may be preferentially perturbed during pulmonary fibrosis.

Effects of inhaled ammonium sulfate on benzo[a]pyrene carcinogenesis.

The effect of inhaled ammonium sulfate on benzo[a]pyrene carcinogenesis in the lungs of Syrian golden hamsters was studied. Exposure to ammonium sulfate at an airborne concentration 20 times average United States ambient levels resulted in a significant depression (p less than 0.05) of benzo[a]pyrene carcinogenesis in the first 6 mo of the study. However, at 2 yr, the termination of the study, there were no differences in cancer incidence between groups receiving benzo[a]pyrene and benzo[a]pyrene plus ammonium sulfate. In addition, at the concentration studied, inhaled ammonium sulfate did not significantly increase the incidence or severity of pneumonitis or pulmonary fibrosis in the hamster. However, this inhalation did increase the incidence of emphysema but not the severity. The decreased incidence of cancer during the first 6 mo of this study in animals receiving both benzo[a]pyrene and ammonium sulfate suggests that interaction between sulfate and benzo[a]pyrene does occur, but is insufficient to afford long-term protection against the development of cancer. No enhancement of carcinogenesis by benzo[a]pyrene occurs in the presence of inhaled sulfate.

Behavior of magnetic particles in hamster lungs: estimates of clearance and cytoplasmic motility.

Ferrimagnetic particles suspended in saline were instilled intratracheally into the lungs of Syrian golden hamsters. The particles were magnetized and aligned by applying an external magnetic field. Upon removal of the external field, the particles produced a remanent magnetic field from the lungs which decayed due to random misalignment of the particles (relaxation). Magnetization and relaxation measurements were performed immediately after instillation, then repeatedly during the first 24 h, and finally at intervals of several days up to 30 days after the instillation. The size of the initial remanent magnetic field immediately following each external magnetization is a measure of the amount of iron oxide in the lungs. It decreased with time, reflecting particle clearance. The rate of relaxation increased steeply during the first 12 h after the instillation and decreased slowly between the 5th and 30th day. Changes in the location of particles from extracellular to intracellular sites and movements from ectoplasmic to endoplasmic sites within cells may be responsible for the observed changes in relaxation rates with time.

Uptake of latex particles by macrophages: characterization using flow cytometry.

Flow cytometry can be used to characterize the uptake of particles by small samples of pulmonary macrophages both quickly and accurately. We found that there was a linear relationship between the number of fluorescent latex particles and the fluorescent intensity associated with each cell up to 47 particles/cell. Macrophages lavaged from the lungs of Syrian golden hamsters were metabolically and functionally stable for 3 h of incubation; the average intracellular concentrations of Na+ and K+ were 29 +/- 2 and 158 +/- 6 mM, respectively. Particle uptake was significantly inhibited by removing divalent cations with ethylenediaminetetraacetic acid and lowering the incubation temperature (37 degrees C) to 24 and 3 degrees C. The cell-to-cell variability in the uptake of particles was greater than the Poisson distribution would predict based on the mean number of particles per cell.

Induction of Lipid Droplets in Fibroblasts of the Hamster Lung by a Diet High in Vitamin A

To investigate the effect of vitamin A on the occurrence of lipid droplets in alveolar fibroblasts, light and electron microscopic studies were carried out on the lungs of Syrian golden hamsters fed on diets with a low, normal, or high vitamin A content for a period of 17 weeks. The feeding of the high vitamin A diet resulted in a considerable increase in size and number of lipid droplets in alveolar fibroblasts. These droplets also occurred in fibroblasts of the visceral pleura, and in fibroblasts of connective tissue around bronchioli and larger blood vessels. Most of the lipid-laden fibroblasts could be recognized as myofibroblasts. The significance of the droplets for the deposition of interstitial materials or for the synthesis of prostaglandins is discussed.

Toxic and transforming effects of cigarette smoke condensates in fetal Syrian golden hamster lung cell cultures

The cytotoxic and transforming effects of cigarette smoke condensates (CSCs) and benzo(a)pyrene (BaP) were compared using cultures of fetal Syrian golden hamster lung cells (FSHL). The dose-response patterns differed completely between CSCs and BaP. The amount of BaP alone failed to explain the transforming activity of CSCs.

Comparison of toxic and transforming effects of ten environment‐related polycyclic hydrocarbons, including benzo [a] pyrene, on Fetal Hamster Lung Cell Cultures

AbstractStandardized procedures were developed to compare quantitatively the toxic and transforming effects of ten environment‐related polycyclic aromatic hydrocarbons (PAH's) using early passage mixed cultures of fetal Syrian golden hamster lung cells. Evaluation of toxicity was attempted by applying a linear regression to the relative number of surviving colonies. Transformation data were fitted satisfactorily to the one‐hit model. Used as a monitor compound in each subexperiment, benzo[a]pyrene (BaP) showed dose‐related colony reduction (0.01–0.5 μg ml−1). Benzo[b]fluoranthene (BbF), indeno[1,2,3‐cd]pyrene (IcdP), benzo[k]fluoranthene (BkF) and benzo[e]pyrene (BeP) were generally colony enhancing at dose ranges up to 1.0 μg ml−1. Fluoranthene (FLT), pyrene (PYR), chrysene (CHR), benzo[ghi]perylene (BghiPL) and benz[a]anthracene (BaA) were neither stimulatory nor inhibitory to colony formation. For transformation induction, BaP was over 14‐fold more effective than BbF, IcdP or BaA. The other PAH's showed no transformation.

Neuroepithelial bodies (NEB) and solitary endocrine cells in the hamster lung.

Periodic acid-Schiff (PAS)-positive small-granule endocrine cells identified in serial plastic sections through 95% of the infracardiac lobe of a Syrian golden hamster lung were marked on a 70X cardboard reconstruction of the airways and recorded for computerized analysis as described in the preceding paper in this volume [1]. When airways were subdivided into thirds and the resulting small pieces of airway surface were analyzed statistically, endocrine cell loci appeared to be randomly distributed in the epithelial layer. Nevertheless, loci were absent from only 10 of 209 unit airways, far short of the 28 predicted from a random distribution. Solitary endocrine cells accounted for 37% of 980 loci but only 6% of all endocrine cells; 63% of loci consisted of endocrine cell clusters, broadly defined as "neuroepithelial bodies," of from 2 to 107 cells. Although small groups were preponderant, no significant discontinuity occurred in the size distribution between 1 and 56 cells per locus. Neuroepithelial bodies predominated everywhere, but especially in the lobar bronchus and at bronchioloalveolar portals, where cuboidal bronchiolar epithelium gives way to the attenuated epithelium of the respiratory zone; 45% were related to peribronchial smooth muscle and 18% to pulmonary capillaries. Only 3.3% of solitary cells were related to capillaries, 29% to airway muscle; 68% occurred in muscle-free regions of the airway, associated preferentially with ciliated epithelial cells. We conclude that endocrine cells are distributed in such a way that virtually all airways have at least one locus. This nonrandomness suggests that these cells are in fact important to normal lung function. Furthermore, because solitary endocrine cells and "neuroepithelial" cell clusters have different intrapulmonary distributions, they may well have distinct identities and functions.

Sensitivity of Syrian golden hamster fetal lung cells to benzo[ a]pyrene and other polycyclic hydrocarbons in vitro

Dose responses were compared of cultured fetal Syrian golden hamster lung cells (FSHL) to the toxic and transforming effects of benzo[ a]pyrene (B[ a]P), benzo[ b]fluoranthene (B[ b]F), benz[ a]anthracene (B[ a]a) indeno[1,2,3- c,d]pyrene (I[ c,d]P), benzo[ k]fluoranthene (B[ k]F) and benzo[ e]pyrene (B[ e]P). Effort was first given to standardising the techniques for evaluating B[ e]P dose-responses. These polycyclic aromatic hydrocarbons (PAH) were then tested at concentrations of up to 1 μg/ml, and only B[ a]P showed clear cytotoxicity. The transforming effects of B[ b]F, B[ a]A and I[ c,d]P at 1 μg/ml appeared comparable to those of B[ a]P at 0.05 μg/ml.