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With some exceptions, global policymakers have recommended against the use of existing monoclonal antibodies in COVID-19 due to loss of neutralization of newer variants. The purpose of this study was to investigate the impact of Ronapreve on compartmental viral replication using paradigms for susceptible and insusceptible variants. Virological efficacy and impact on pathogenicity was assessed in K18-hACE2 mice inoculated with either the Delta or BA.1 Omicron variants. Ronapreve reduced sub-genomic viral RNA levels in lung and nasal turbinate, 4 and 6 days post-infection, for the Delta variant but not the Omicron variant. It also blocked brain infection, which is seen with high frequency in K18-hACE2 mice after Delta variant infection. At day 6, the inflammatory response to lung infection with the Delta variant was altered to a multifocal granulomatous inflammation in which the virus appeared to be confined. The current study provides evidence of an altered tissue response to SARS-CoV-2 after treatment with a monoclonal antibody combination that retains neutralization activity. These data demonstrate that experimental designs that reflect treatment use cases are achievable in animal models for monoclonal antibodies. Extreme caution should be taken when interpreting prophylactic experimental designs that may not be representative of treatment.IMPORTANCEFollowing the emergence of the SARS-CoV-2 Omicron variant, the WHO recommended against the use of Ronapreve in its COVID-19 treatment guidelines due to a lack of efficacy based on current pharmacokinetic-pharmacodynamic understanding. However, the continued use of Ronapreve, specifically in vulnerable patients, was advocated by some based on in vitro neutralization data. Here, the virological efficacy of Ronapreve was demonstrated in both the lung and brain compartments using Delta as a paradigm for a susceptible variant. Conversely, a lack of virological efficacy was demonstrated for the Omicron variant. Comparable concentrations of both monoclonal antibodies were observed in the plasma of Delta- and Omicron-infected mice. This study made use of a reliable murine model for SARS-CoV-2 infection, an experimental design reflective of treatment, and demonstrated the utility of this approach when assessing the effectiveness of monoclonal antibodies., Competing Interests: A.O. and S.R. are directors of Tandem Nano Ltd. and co-inventors of patents relating to drug delivery. A.O. has been a co-investigator on funding received by the University of Liverpool from ViiV Healthcare and Gilead Sciences unrelated to COVID-19 in the past 3 years. A.O. has received personal fees from Gilead and Assembly Biosciences in the past 3 years unrelated to COVID-19. A.O. was a member of the Trial Management Group for the AGILE phase I/II platform trial until January 2023 and AGILE received funding from Ridgeback and GSK in the past 3 years for which A.O. was not a co-investigator. S.R. has received research funding from ViiV and AstraZeneca, and consultancy from Gilead not related to the current paper. No other conflicts are declared by the authors.

Objectives: Antiviral interventions are required to complement vaccination programmes and reduce the global burden of COVID-19. Prior to initiation of large-scale clinical trials, robust preclinical data to support candidate plausibility are required. This work sought to further investigate the putative antiviral activity of probenecid against SARS-CoV-2., Methods: Vero E6 cells were preincubated with probenecid, or control media for 2 h before infection (SARS-CoV-2/Human/Liverpool/REMRQ0001/2020). Probenecid or control media was reapplied, plates reincubated and cytopathic activity quantified by spectrophotometry after 48 h. In vitro human airway epithelial cell (HAEC) assays were performed for probenecid against SARS-CoV-2-VoC-B.1.1.7 (hCoV-19/Belgium/rega-12211513/2020; EPI_ISL_791333, 2020-12-21) using an optimized cell model for antiviral testing. Syrian golden hamsters were intranasally inoculated (SARS-CoV-2 Delta B.1.617.2) 24 h prior to treatment with probenecid or vehicle for four twice-daily doses., Results: No observable antiviral activity for probenecid was evident in Vero E6 or HAEC assays. No reduction in total or subgenomic RNA was observed in terminal lung samples (P > 0.05) from hamsters. Body weight of uninfected hamsters remained stable whereas both probenecid- and vehicle-treated infected hamsters lost body weight (P > 0.5)., Conclusions: These data do not support probenecid as a SARS-CoV-2 antiviral drug., (© The Author(s) 2023. Published by Oxford University Press on behalf of British Society for Antimicrobial Chemotherapy.)

Background: It is not understood why some pulmonary fibroses such as cryptogenic organizing pneumonia (COP) respond well to treatment, while others like usual interstitial pneumonia (UIP) do not. Increased understanding of the structure and function of the matrix in this area is critical to improving our understanding of the biology of these diseases and developing novel therapies. The objectives herein are to provide new insights into the underlying collagen- and matrix-related biological mechanisms driving COP versus UIP., Methods: Two-photon second harmonic generation (SHG) and excitation fluorescence microscopies were used to interrogate and quantify differences between intrinsic fibrillar collagen and elastin matrix signals in healthy, COP, and UIP lung., Results: Collagen microstructure was different in UIP versus healthy lung, but not in COP versus healthy, as indicated by the ratio of forward-to-backward propagating SHG signal (FSHG/BSHG). This collagen microstructure as assessed by FSHG/BSHG was also different in areas with preserved alveolar architecture adjacent to UIP fibroblastic foci or honeycomb areas versus healthy lung. Fibrosis was evidenced by increased col1 and col3 content in COP and UIP versus healthy, with highest col1:col3 ratio in UIP. Evidence of elastin breakdown (i.e. reduced mature elastin fiber content), and increased collagen:mature elastin ratios, were seen in COP and UIP versus healthy., Conclusions: Fibrillar collagen's subresolution structure (i.e. "microstructure") is altered in UIP versus COP and healthy lung, which may provide novel insights into the biological reasons why unlike COP, UIP is resistant to therapies, and demonstrates the ability of SHG microscopy to potentially distinguish treatable versus intractable pulmonary fibroses.

Ovine pulmonary adenocarcinoma (OPA) is a contagious tumour caused by infection of sheep with Jaagsiekte sheep retrovirus. Two forms of OPA have been identified, classical and atypical, which can be distinguished clinically and pathologically. Most notably classical OPA is progressive until death, while atypical OPA remains subclinical. In the present study the local immune responses in the lungs of cases of atypical OPA were compared with those from classical cases by immunohistochemistry using a panel of mouse anti-sheep mAbs. Distinct differences in the distribution of immune cell subsets in the two forms of OPA were observed. In particular there was an intratumoural influx of T cell subsets and MHC Class II expression on the tumour cells in atypical OPA, neither of which was seen in classical OPA. It is possible that these differences may contribute, at least in part, to determining the progressive course of classical OPA compared with the subclinical nature of atypical OPA., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Bone marrow (stem) cells can differentiate into cells in multiple tissues, including lung. Conversely, there are reports that cells of nonhematopoietic tissues (brain, muscle) can give rise to lymphohematopoietic cells. Here we show that the lung contains cells capable of giving rise to lymphohematopoietic cells when transplanted to irradiated recipients. Whole lung cell suspensions, lung side population (SP) cells, and CD45(+/-) lung cells obtained from male transgenic enhanced green fluorescent protein-expressing mice were transplanted intravenously to total body irradiated female mice. Green fluorescent cells were recovered from the circulation and phenotyped for their expression of lymphohematopoietic markers (CD3, CD4, CD8, B220, Gr-1, and Mac-1). Lung SP cells were composed of heterogeneous populations and had less ability to give rise to lymphohematopoietic cells than did bone marrow SP cells. Furthermore, the ability of cells from the lung of aged mice to generate lymphohematopoietic progeny was equivalent to that of cells from young mice. Cells from lung with radioprotective and lymphohematopoietic reconstituting abilities were CD45(+). CD45(+) cells in the lung cells have lymphohematopoietic stem/progenitor cell characteristics, and this has implications for cell or gene therapy applications.

Background: BM cells have been shown to give rise to progeny of various cell lineages, including cells in lung and liver. This investigation evaluated whether purified BM mononuclear cells and side population (SP) cells that have hematopoietic stem-cell activity also had this property; whether a TBI preparative regimen was necessary for engraftment; and where BM-derived cells were engrafted., Methods: Either 1-3 million BM mononuclear cells or 2000 BM SP cells from transgenic enhanced green fluorescent protein-expressing (EGFP) mice were transplanted i.v. to unirradiated or 7-9.5 Gy irradiated recipients., Results: Flow cytometric analysis showed that lung cells (mean 45%, range 4-70%) and liver cells (mean 4%, range 0.4-8.3%) from irradiated, but not unirradiated recipients, were EGFP donor-derived. Similar results were obtained transplanting BM mononuclear cells or SP cells. Morphologically, donor-derived cells in the lung were primarily monocytes and macrophages. Additionally, lung fibroblasts and Type I, but not Type II, alveolar cells and rare cells in the bronchial epithelium were donor BM derived. In the liver, Kupffer cells, inflammatory cells and small clusters of hepatocytes, but not bile duct cells, were donor-derived., Discussion: BM mononuclear and SP cells generated progeny in some compartments of the lung and liver, but only in TBI recipients. Stem cells in BM can contribute to repair of tissue injury in some compartments, but not to the same extent in the lung and liver.

Sheep pulmonary adenomatosis (SPA) is a naturally occurring contagious lung tumour of sheep which has been associated aetiologically with a type D- and B-related retrovirus (Jaagsiekte retrovirus; JSRV). To improve understanding of the aetio-pathogenesis of SPA, the distribution and the sites of JSRV replication in sheep with naturally or experimentally induced SPA or in unaffected controls were identified. New immunological reagents were produced and a blocking enzyme-linked immunosorbent assay (B-ELISA) and an immunohistochemical technique for the detection of JSRV major capsid protein at the tissue and cellular levels were developed. JSRV was detected only in the respiratory tract of sheep affected by pulmonary adenomatosis and specifically in the transformed epithelial cells of the alveoli of SPA-affected sheep.

In anesthetized dogs studied both supine and prone, the electromyogram (EMG) of the diaphragm recorded directly from three portions of the diaphragm (crural, anterior, and costal) was compared with simultaneous recordings of the diaphragmatic EMG recorded from 10 sites in the esophagus and stomach. Effects upon the EMG of lung volume change and esophageal electrode position change were determined during bilateral supramaximal tetanic phrenic stimulation with airway occluded. Lung volume change had little effect upon the directly recorded EMG. The effect of lung volume change upon the EMG recorded from the esophagus was somewhat greater and marked change was noted as the esophageal recording site was varied. In supine dogs two sites of maximal signal were observed, one 1 cm above the cardia and the other 4--6 cm below the cardia. In prone dogs a single site for maximal signal was observed 3 cm above the cardia. An electrode site as close to the cardia as possible appears to be optimal from the point of view of variation in signal due to lung volume change and due to body position change. Gastric balloon stabilization is recommended. Proximity of the electrode and posterior gastric wall to the diaphragmatic crura may explain the maximal EMG signal recorded below the cardia.

The infective agent of jaagsiekte was shown to be present in the fluid which accumulates in the respiratory tract of sheep during the terminal stages of the disease. The fluid also contained reverse transcriptase (RT) activity which showed a clear preference for a ribonucleic acid synthetic template over the corresponding deoxyribonucleic acid template and which utilised the RT specific template/primer poly (2'-0-methylcytidylate) oligodeoxyguanylate. This enzyme activity was associated with a particle which had typical retroviral buoyant densities in a range of gradient media.

There is controversy whether routine use of theophylline is warranted in chronic obstructive pulmonary disease (COPD). Effects on spirometry are minimal, and subjective relief is difficult to document. Therefore, we undertook to study the pulmonary effects of theophyline using a functional approach, namely, its effects on the lung mechanics of a standing or walking patient. In a double blind, placebo controlled crossover study, 400 mg of slow-release theophylline (TheoDur) were given twice a day in 13 trials to 10 patients with severe COPD. Lung mechanics were calculated on a breath-by-breath basis by a pulmonary mechanics computer from esophageal pressures and flow. Expired volumes were simultaneously collected. Spirometry was then performed. On the placebo limb, the spirometric response to 0.5 mg terbutaline given subcutaneously was compared with the theophylline response. At a mean serum theophylline level of 12.3 micrograms/ml, theophylline produced in the entire group a mean reduction in the work of breathing done on the lung (WL) of 16% (p less than 0.002) while walking, but a significant effect while standing was not detected. During both walking and standing, the mean reduction in lung resistance (RL) was 15 to 16% (p less than 0.002). Minute volume (VE) was increased at borderline significance, with a mean reduction while walking of 19% in the ratio of WL to VE (p less than 0.002). In the individual patient, changes in WL were influenced more by changes in VE than by changes in RL; in fact, changes in WL correlated best with changes in the product of (VE)2 and RL. Theophylline produced a mean increase of 15% in forced expiratory volume in one second (FEV1) and 16% in forced vital capacity (FVC).(ABSTRACT TRUNCATED AT 250 WORDS)

Measurements of pulmonary function were made on patients within 1 wk, 3 wk, 5 wk, 3 months, and 5 months of spinal cord injury. In patients with functionally complete transection of the cord between segment C5 and C6, the vital capacity was 30% of predicted in the first week after injury. Patients with injuries at C4 had smaller vital capacities. A significant increase in vital capacity occurred within 5 wk of injury with an approximate doubling of vital capacity 3 months after injury. Expiratory flow rates were directly dependent upon vital capacity. There was a high incidence of arterial hypoxemia, in the acute stage, even in patients with adequate ventilatory ability and normocarbia.

An efficient and reproducible technique is described for the isolation of transformed sheep pulmonary adenomatosis cells. It includes three basic steps: prolonged trypsinisation to kill fibroblasts, magnetic removal of macrophages and adherence to remove the rapidly adherent cells. The resultant preparations of lung cells were enriched to 96.6 per cent type 2 pneumocytes.

To define the regional ventilation of the human lung, subjects were monitored with a gamma camera, while breathing air labeled with radioactive 81mKr which is being continuously eluted from a generator. The information observed is displayed as a reiterated multiframe moving picture. This is accomplished by storing the information from multiple breaths, selecting acceptable cycles (determined by the duration of inspiration and expiration and the amplitude to extremes of the cycles), and normalizing the usable information and converting the summed data into sequential frames, which are repeatedly shown on a video screen from computer memory.

Seven lambs were challenged with an aerosol of OA4 virus. Four developed mild pulmonary oedema and a sparse accumulation of mononuclear cells around blood vessels similar to results reported previously. In addition, three had focal hepatic necrosis and one of these three had a non-suppurative occlusive cholangitis and portal tract lymphangitis and thrombosis. Basophilic intranuclear IB's were identified in many necrotic hepatocytes, in lymphatic endothelial cells involved in thrombi but in only one bile duct epithelial cell. Ultrastructurally, virions with the characteristics of adenovirus were seen in hepatocytes. Although only one IB was seen in the affected biliary system there was no evidence that it was caused by an agent other than OA4. Virus was considered to infect the respiratory system directly, be swallowed to infect the alimentary system and carried, possibly within cells, via a haematogeneous and/or lymphogenous route to the liver. The effect of naturally acquired virus on sheep is unknown but it is possible that it may predispose the host to more serious infections.

Two retrovirus-associated pulmonary diseases of sheep [ovine pulmonary carcinoma (OPC); sheep pulmonary adenomatosis], a bronchoalveolar carcinoma, and lymphoid interstitial pneumonia (LIP) were induced simultaneously in 9 of 9 neonatal lambs. The lambs were killed 8-28 weeks after intratracheal injection of lung tumor homogenate or lung fluid derived from sheep with naturally occurring OPC and ovine lentivirus (OvLV) infection. The inoculated lambs developed multifocal neoplasms of alveolar type II cells or nonciliated bronchiolar epithelial cells, LIP, and pulmonary lymph node hyperplasia, and all produced antibody to OvLV. OvLV was isolated from 6 to 7 lambs tested, and infectious center assay of pulmonary lavage cells from 3 lambs revealed that approximately 1 in 1,000 pulmonary lavage cells contained infectious lentivirus. Neither contact control lambs nor control lambs that received ultrafiltered lung fluid developed evidence of either disease or of OvLV infection. Lung fluid or tumor tissue of lambs with OPC contained a 26,000-dalton protein that cross-reacted with antiserum to p27 to Mason-Pfizer monkey virus, a type D retrovirus. The fact that no antigenic cross-reaction between OvLV and type D retroviruses has been demonstrated supports the presence of two retroviruses in sheep with OPC. Although the contributions of each agent to oncogenesis in this model are difficult to evaluate, the rapid development of two retrovirus-induced pulmonary diseases in experimentally inoculated lambs suggests an etiologic or pathogenetic synergism between these two members of the family Retroviridae.

Five sheep with ovine pulmonary carcinoma were markedly dyspneic and had sporadic coughing; two had copious watery nasal exudate. In four, lesions consisted of multifocal nodules of neoplastic cuboidal epithelial cells in acinar or papillary patterns. Electron microscopically, cells had microvilli, tight junctions, and cytoplasmic lamellar bodies typical of alveolar type II cells. One sheep had a single lung tumor of nonciliated bronchiolar epithelial cells. Vacuolated alveolar macrophages surrounded adenomatous foci. One sheep had a metastatic lesion in the caudal mediastinal lymph node. All sheep had histologic lesions of lymphoid interstitial pneumonia (LIP, ovine progressive pneumonia) consisting of peribronchiolar and interstitial lymphoid hyperplasia, and fibromuscular proliferation; all had serum precipitating antibodies to ovine lentivirus. Lung fluids or tumor homogenates contained a 26-kd peptide that crossreacted with a primate-derived type D retrovirus as detected by immunoblotting or interspecies competition radioimmunoassay. Ovine lentivirus was isolated from concentrated lung fluids or tumor tissues of four sheep tested and from tumor cell DNA of one animal transfected into ovine muscle cells. These studies document the presence of type D-related retrovirus antigen in ovine pulmonary carcinoma (OPC) in the United States and indicate that lentivirus-induced LIP is a lesion frequently associated with this disease.