scholarly journals Identification and phenotype of MAIT cells in cattle and their response to bacterial infections

2020 ◽  
Author(s):  
Matthew D. Edmans ◽  
Timothy K. Connelley ◽  
Siddharth Jayaraman ◽  
Christina Vrettou ◽  
Martin Vordermeier ◽  
...  
Keyword(s):  
Immune Response ◽  
Peripheral Blood ◽  
Bacterial Infections ◽  
Large Animal Model ◽  
Effector Memory ◽  
Large Animal ◽  
Riboflavin Biosynthesis ◽  
Mait Cells ◽  
Zoonotic Infections ◽  
Α Chain

AbstractMucosal-associated invariant T (MAIT) cells are a population of innate-like T cells that utilise a semi-invariant T cell receptor (TCR) α chain and are restricted by the highly conserved antigen presenting molecule MR1. MR1 presents microbial riboflavin biosynthesis derived metabolites produced by bacteria and fungi. Consistent with their ability to sense ligands derived from bacterial sources, MAIT cells have been associated with the immune response to a variety of bacterial infections, such as Mycobacterium spp., Salmonella spp. and Escherichia coli. To date, MAIT cells have been studied in humans, non-human primates and mice. However, they have only been putatively identified in cattle by PCR based methods; no phenotypic or functional analyses have been performed. Here, we identified a MAIT cell population in cattle utilising MR1 tetramers and high-throughput TCR sequencing. Phenotypic analysis of cattle MAIT cells revealed features highly analogous to those of MAIT cells in humans and mice, including expression of an orthologous TRAV1-TRAJ33 TCR α chain, an effector memory phenotype irrespective of tissue localisation, and expression of the transcription factors PLZF and EOMES. We determined the frequency of MAIT cells in peripheral blood and multiple tissues, finding that cattle MAIT cells are enriched in mucosal tissues as well as in the mesenteric lymph node. Cattle MAIT cells were responsive to stimulation by 5-OP-RU and riboflavin biosynthesis competent bacteria in vitro. Furthermore, MAIT cells in milk increased in frequency in cows with mastitis. Following challenge with virulent Mycobacterium bovis, a causative agent of bovine tuberculosis and a zoonosis, peripheral blood MAIT cells expressed higher levels of perforin. Thus MAIT cells are implicated in the immune response to two major bacterial infections in cattle. These data suggest that MAIT cells are functionally highly conserved and that cattle are an excellent large animal model to study the role of MAIT cells in important zoonotic infections.

scholarly journals Identification and Phenotype of MAIT Cells in Cattle and Their Response to Bacterial Infections

2021 ◽  
Vol 12 ◽  
Author(s):  
Matthew D. Edmans ◽  
Timothy K. Connelley ◽  
Siddharth Jayaraman ◽  
Christina Vrettou ◽  
Martin Vordermeier ◽  
...  
Keyword(s):  
Immune Response ◽  
Peripheral Blood ◽  
Bacterial Infections ◽  
Large Animal Model ◽  
Effector Memory ◽  
Large Animal ◽  
Riboflavin Biosynthesis ◽  
Mait Cells ◽  
Zoonotic Infections ◽  
Α Chain

Mucosal-associated invariant T (MAIT) cells are a population of innate-like T cells that utilize a semi-invariant T cell receptor (TCR) α chain and are restricted by the highly conserved antigen presenting molecule MR1. MR1 presents microbial riboflavin biosynthesis derived metabolites produced by bacteria and fungi. Consistent with their ability to sense ligands derived from bacterial sources, MAIT cells have been associated with the immune response to a variety of bacterial infections, such as Mycobacterium spp., Salmonella spp. and Escherichia coli. To date, MAIT cells have been studied in humans, non-human primates and mice. However, they have only been putatively identified in cattle by PCR based methods; no phenotypic or functional analyses have been performed. Here, we identified a MAIT cell population in cattle utilizing MR1 tetramers and high-throughput TCR sequencing. Phenotypic analysis of cattle MAIT cells revealed features highly analogous to those of MAIT cells in humans and mice, including expression of an orthologous TRAV1-TRAJ33 TCR α chain, an effector memory phenotype irrespective of tissue localization, and expression of the transcription factors PLZF and EOMES. We determined the frequency of MAIT cells in peripheral blood and multiple tissues, finding that cattle MAIT cells are enriched in mucosal tissues as well as in the mesenteric lymph node. Cattle MAIT cells were responsive to stimulation by 5-OP-RU and riboflavin biosynthesis competent bacteria in vitro. Furthermore, MAIT cells in milk increased in frequency in cows with mastitis. Following challenge with virulent Mycobacterium bovis, a causative agent of bovine tuberculosis and a zoonosis, peripheral blood MAIT cells expressed higher levels of perforin. Thus, MAIT cells are implicated in the immune response to two major bacterial infections in cattle. These data suggest that MAIT cells are functionally highly conserved and that cattle are an excellent large animal model to study the role of MAIT cells in important zoonotic infections.

scholarly journals Distinct cellular immune profiles in the airways and blood of critically ill patients with COVID-19

2020 ◽  
Author(s):  
Anno Saris ◽  
Tom D.Y. Reijnders ◽  
Esther J. Nossent ◽  
Alex R. Schuurman ◽  
Jan Verhoeff ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cells ◽  
T Cell ◽  
T Cell Activation ◽  
Peripheral Blood ◽  
Cell Activation ◽  
Effector Memory ◽  
Activated T Cells ◽  
Immune Profile ◽  
Prolonged Icu Stay

AbstractOur understanding of the coronavirus disease-19 (COVID-19) immune response is almost exclusively derived from studies that examined blood. To gain insight in the pulmonary immune response we analysed BALF samples and paired blood samples from 17 severe COVID-19 patients. Macrophages and T cells were the most abundant cells in BALF. In the lungs, both CD4 and CD8 T cells were predominantly effector memory cells and expressed higher levels of the exhaustion marker PD-1 than in peripheral blood. Prolonged ICU stay associated with a reduced proportion of activated T cells in peripheral blood and even more so in BALF. T cell activation in blood, but not in BALF, was higher in fatal COVID-19 cases. Increased levels of inflammatory mediators were more pronounced in BALF than in plasma. In conclusion, the bronchoalveolar immune response in COVID-19 has a unique local profile that strongly differs from the immune profile in peripheral blood.SummaryThe bronchoalveolar immune response in severe COVID-19 strongly differs from the peripheral blood immune profile. Fatal COVID-19 associated with T cell activation blood, but not in BALF.

Posttransplantation lymphoproliferative disease in miniature swine after allogeneic hematopoietic cell transplantation: similarity to human PTLD and association with a porcine gammaherpesvirus

Blood ◽  
2001 ◽  
Vol 97 (5) ◽  
pp. 1467-1473 ◽  
Author(s):  
Christene A. Huang ◽  
Yasushi Fuchimoto ◽  
Zachary L. Gleit ◽  
Thomas Ericsson ◽  
Adam Griesemer ◽  
...  
Keyword(s):  
Animal Model ◽  
Stem Cell ◽  
Stem Cell Transplantation ◽  
Cell Transplantation ◽  
Peripheral Blood ◽  
Lymphoproliferative Disease ◽  
Large Animal Model ◽  
Large Animal ◽  
Miniature Swine ◽  
Hematopoietic Stem

Posttransplantation lymphoproliferative disease (PTLD) is a major complication of current clinical transplantation regimens. The lack of a reproducible large-animal model of PTLD has limited progress in understanding the pathogenesis of and in developing therapy for this clinically important disease. This study found a high incidence of PTLD in miniature swine undergoing allogeneic hematopoietic stem cell transplantation and characterized this disease in swine. Two days before allogeneic peripheral blood stem cell transplantation, miniature swine were conditioned with thymic irradiation and in vivo T-cell depletion. Animals received cyclosporine daily beginning 1 day before transplantation and continuing for 30 to 60 days. Flow cytometry and histologic examination were performed to determine the cell type involved in lymphoproliferation. Polymerase chain reaction was developed to detect and determine the level of porcine gammaherpesvirus in involved lymph node tissue. PTLD in swine is morphologically and histologically similar to that observed in human allograft recipients. Nine of 21 animals developed a B-cell lymphoproliferation involving peripheral blood (9 of 9), tonsils, and lymph nodes (7 of 9) from 21 to 48 days after transplantation. Six of 9 animals died of PTLD and 3 of 9 recovered after reduction of immunosuppression. A novel porcine gammaherpesvirus was identified in involved tissues. Miniature swine provide a genetically defined large-animal model of PTLD with many characteristics similar to human PTLD. The availability of this reproducible large-animal model of PTLD may facilitate the development and testing of diagnostic and therapeutic approaches for prevention or treatment of PTLD in the clinical setting.

Therapeutic Levels of Gene Corrected Neutrophils Following Foamy Viral Vector Mediated Gene Transfer in a Canine Model of Leukocyte Adhesion Deficiency.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 458-458
Author(s):  
Thomas R. Bauer ◽  
James M. Allen ◽  
Laura M. Tuschong ◽  
Erik M. Olson ◽  
Tanya Burkholder ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Bacterial Infections ◽  
Viral Vectors ◽  
Viral Vector ◽  
Leukocyte Adhesion ◽  
Large Animal Model ◽  
Large Animal ◽  
Gene Therapy Approach ◽  
Cd34 Cells ◽  
Post Infusion

Abstract Children with the genetic immunodeficiency disease leukocyte adhesion deficiency (LAD) experience life-threatening bacterial infections due to the inability of their leukocytes to adhere and migrate to sites of infection. Heterogenous molecular defects in the leukocyte integrin CD18 molecule are responsible for LAD. The canine form of LAD, known as CLAD, represents a disease-specific, large animal model in which affected animals typically die within the first few months of life. We evaluated a gene therapy approach in treating CLAD using foamy viral vectors. Foamy viral vectors were selected because of their efficacy of marking in several animal models, including human CD34+ cells in NOD-SCID mice, and because foamy viral vectors may be less likely to cause insertional mutagenesis than conventional Moloney murine leukemia virus (MLV) based vectors. In contrast to MLV, no case of leukemia or disease has been reported in animals or humans as a consequence of infection by foamy viruses. Each of three CLAD dogs received a single infusion of autologous, foamy viral vector-transduced CD34+ cells following non-myeloablative conditioning with 200-cGy total body irradiation (TBI). CLAD CD34+ cells were transduced via a short-duration (14–20 hr) exposure to foamy viral vector ΔΦ Mscv-CD18 (2-3 MOI) and 50 ng/mL growth factors cG-CSF, cSCF, and hFlt3-L, on 10 μg/cm2 Retronectin™. Estimated transduction of the CLAD CD34+ cells, using a sample of transduced cells incubated in vitro for 3 additional days, ranged from 13.7 to 24.6% by flow cytometric detection of CD18 expression. The total infused cell doses ranged from 1.7 to 2.3 x 106 CD34+ cells / kg. Two of the three CLAD dogs are alive and well 2 and 6 months post-infusion. By 3 weeks post infusion, both dogs displayed approximately 1.2–1.4% CD18+ neutrophils in the peripheral blood. This level has increased progressively in each animal, to 2.1% CD18+ neutrophils by 6 months in the first dog and to 2.3% by 2 months in the second dog following infusion. Both animals have had resolution or pre-emption of severe CLAD disease when compared to untreated controls, who were euthanized by 6 months due to refractory bacterial infections. The third dog died 6 days after infusion from intussusception arising as a complication from the TBI. These results represent the first report of successful gene therapy in a disease-specific, large-animal model using foamy viral vectors to reverse the disease phenotype. Foamy viral vectors offer potential advantages over MLV-based vectors, including the ability to transduce non-dividing cells and the safety of a non-pathogenic virus backbone. The simplified, short-duration transduction regimen used with foamy viral vectors, coupled with a clinically applicable, non-myeloablative regimen of 200 cGy TBI, produced levels of CD18-gene corrected leukocytes in the peripheral blood that reversed or prevented the disease phenotype in CLAD. These studies indicate that foamy viral vectors may represent a new therapeutic gene therapy approach for the treatment of children with LAD. Long-term follow-up of these animals will be required to establish the efficacy and safety of this vector.

scholarly journals Specific-Pathogen-Free Pigs as an Animal Model for Studying Chlamydia trachomatis Genital Infection

2005 ◽  
Vol 73 (12) ◽  
pp. 8317-8321 ◽  
Author(s):  
Daisy Vanrompay ◽  
Thi Q. T. Hoang ◽  
Liselotte De Vos ◽  
Kristel Verminnen ◽  
Taher Harkinezhad ◽  
...  
Keyword(s):  
Immune Response ◽  
Animal Model ◽  
Chlamydia Trachomatis ◽  
Immunoglobulin M ◽  
Large Animal Model ◽  
Genital Infection ◽  
Large Animal ◽  
Specific Pathogen Free ◽  
Humoral Immune ◽  
Specific Pathogen

ABSTRACT The purpose of the present study was to evaluate pigs as a large-animal model for female genital infection with two Chlamydia trachomatis human serovar E strains. Sixteen-week-old specific-pathogen-free female pigs (gilts) were intravaginally infected with the trachoma type E reference strain Bour or the urogenital serovar E strain 468. Several conclusions can be drawn from our findings on the pathogenicity of a primary C. trachomatis genital infection in gilts. First of all, we demonstrated that the serovar E strains Bour and 468 could ascend in the genital tract of gilts. The serovar E strains could replicate in the superficial columnar cervical epithelium and in the superficial epithelial layer of the uterus, which are known to be the specific target sites for a C. trachomatis genital infection in women. Second, inflammation and pathology occurred at the replication sites. Third, the organisms could trigger a humoral immune response, as demonstrated by the presence of immunoglobulin M (IgM), IgG, and IgA in both serum and genital secretion samples. Our findings imply that the pig model might be useful for studying the pathology, pathogenesis, and immune response to a C. trachomatis infection of the genital system.

scholarly journals Hemophilia A Dogs Tolerant to Human Factor VIII Provide a Unique Model to Determine Efficacy and Safety of AAV Delivery of Novel Factor VIII Variants

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 3628-3628
Author(s):  
Giang N. Nguyen ◽  
Lauren E. Wimsey ◽  
Elizabeth P. Merricks ◽  
Katherine P. Ponder ◽  
Timothy C. Nichols ◽  
...  
Keyword(s):  
Immune Response ◽  
Animal Model ◽  
Factor Viii ◽  
Human Factor ◽  
Hemophilia A ◽  
Specific Activity ◽  
Large Animal Model ◽  
Large Animal ◽  
Efficacy And Safety ◽  
Therapy Studies

The hemophilia dog models are valuable for evaluating the efficacy of novel hemophilia therapeutics. The hemophilia B dog was predictive of the therapeutic dose of adeno-associated viral (AAV) vector delivery of human factor IX in clinical trials. In addition, it provides an opportunity to study the long-term efficacy and safety after AAV administration. However, there are several challenges in using the hemophilia A (HA) dog model for gene therapy studies. First, canine factor VIII (cFVIII) has higher specific activity and increased rate of secretion compared to human FVIII (hFVIII). This significant difference between cFVIII and hFVIII prevents the use of a species-specific transgene to predict the efficacy of AAV-hFVIII. Second, the HA dogs are immunocompetent and develop an immune response to the xenoprotein, hFVIII, that precludes the ability to measure transgene expression. Therefore, in order to employ this valuable model for gene therapy studies, we generated a unique cohort of HA dogs that are tolerant to B-domain deleted (BDD) hFVIII. We hypothesized that tolerizing dogs to hFVIII will (1) permit accurate evaluation of hFVIII expression and thus predict the therapeutic vector dose and (2) allow the evaluation of the potential immune response to a novel hFVIII variant. To tolerize the dogs to hFVIII, neonatal HA dogs were treated with a retrovirus (RV-hAAT-hFVIII-BDD-WPRE, 3x109 TU/kg) (n=5) on day 2 of life. The hFVIII expression was between 0.3%-6% at 4 weeks after RV delivery and plateaued after 6 months to 0.8% (S28), 0.3% (S29), 0% (V06), 1.5% (V26) and 1.7% (V27) based on Coatest assay. To determine if the dogs were tolerant to hFVIII-BDD, the dogs were challenged with hFVIII-BDD protein at 5-6 months post-RV administration (Xyntha, 25IU/kg per wk x 6 wks, I.V.). Anti-hFVIII antibodies were monitored closely throughout the challenge and up to 8 weeks after the last challenge. In 4 out of 5 dogs, no anti-hFVIII immune response was observed based on IgG1, IgG2, total IgG or Bethesda titer. In contrast, naïve HA dogs (n=2) developed high level anti-hFVIII IgG2 (1.2-3.2 μg/mL), total IgG (3.4-5.0 μg/mL), and Bethesda titer (4.1-67.8 BU/mL) after the same challenge regimen. Interestingly, the hFVIII activity in one RV-treated dog (V06) was undetectable at 6 months post-RV administration. After the challenge, V06 had anti-hFVIII IgG2 (1.7 μg/mL), total IgG (2.6 μg/mL), and a Bethesda titer (9.5 BU/mL), suggesting that FVIII must be maintained to achieve tolerance. These dogs were used to evaluate the efficacy of AAV serotype 8 (AAV8) delivery of a hFVIII-BDD codon-optimized sequence driven by a hepatocyte promoter, modified transthyretin promoter (TTRm). S29 was delivered AAV8-TTRm-hFVIII-CO (2x1012 vg/kg). Prior to AAV delivery, the levels of hFVIII activity were 0.5-1% from the tolerization with the RV. After AAV administration the hFVIII activity was 3.8% at d168 and 4.7% at d387, resulting in a 4% increase in hFVIII expression. No anti-hFVIII antibodies were detected. The annual bleeding rate (ABR) for S29 post-RV delivery was 5 and after AAV delivery was 0, showing an improvement in the bleeding phenotype in contrast to untreated HA dogs (ABR=13, n=11). A hFVIII-tolerized littermate, S28, was recently treated with a hFVIII variant, AAV8-TTRm-hFVIII-CO-Δ3-SP/DE (2x1012 vg/kg). The hFVIII-Δ3-SP/DE variant has a deletion of the furin site (1645-47) and replaces residues SD at 1657-58 with PE. This variant showed higher specific activity (2-fold) in vitro and increased secretion (4-fold) compared to wild type hFVIII-BDD in the setting of AAV delivery in HA mice. Based on the results in S28, we will determine the dosing of V26 and V27. These studies demonstrate that sustained low level hFVIII expression of 0.2-2% up to 4 years post-retroviral delivery were able to induce and maintain tolerance to hFVIII, while allowing for the subsequent assessment of AAV efficacy. A clinically relevant dose of AAV8-TTRm-hFVIII-CO resulted in therapeutic levels of hFVIII expression while ongoing studies will allow investigation of the efficacy of the hFVIII-BDD variant, Δ3-SP/DE, in the setting of AAV administration in a large animal model. Overall, these studies demonstrate that RV-targeting of hFVIII-BDD expression to the liver in neonatal HA dogs leads to tolerance to this xenoprotein and provide a unique large animal model to evaluate both efficacy as well as potential immunogenicity of novel FVIII variants. Disclosures Sabatino: Spark Therapeutics: Patents & Royalties.

Development and Characterization of a Novel CD34 Monoclonal Antibody That Identifies Sheep Hematopoietic Stem/Progenitor Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4909-4909 ◽  
Author(s):  
Christopher D. Porada ◽  
Dee Harrison-Findik ◽  
Chad Sanada ◽  
Vincent Valiente ◽  
Paul J. Simmons ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Animal Model ◽  
Monoclonal Antibodies ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Large Animal Model ◽  
Large Animal ◽  
Sheep Model ◽  
Cd34 Cells ◽  
Hsc Transplantation

Abstract To date, the cell surface marker CD34 has been the antigen most widely used for identification and isolation of HSC in humans and numerous other species. CD34 is an integral membrane sialomucin present on roughly 1–3% of BM mononuclear cells (MNC) in a normal adult human, yet its precise function remains largely unknown. The CD34+ population is now known to produce durable reconstitution of all blood lineages in both autologous and allogeneic transplantations, providing evidence that CD34 is expressed on some of the most primitive long-term engrafting HSC. We and others have used the fetal sheep model extensively to study the potential and behavior of human HSC. Unfortunately, no reagents exist that allow sheep HSC to be identified or purified, impeding the development of experimental HSC transplantation strategies in this clinically relevant large animal model. We therefore developed monoclonal antibodies (MoAbs) to the ovine homologue of CD34. We PCR cloned and sequenced an 858bp cDNA corresponding to the extracellular domain of the sheep CD34 antigen, which was found to share 92% homology with the bovine CD34 homologue. When aligned to human CD34, the sheep homologue shared 90% homology from nucleotides 1–84, 78% homology from nucleotides 453–858, and exhibited minimal homology between nucleotides 85 and 452. This cDNA was cloned into an expression vector and used to genetically immunize mice and create monoclonal antibodies. One antibody (8D11) was selected for all subsequent studies. Using flow cytometry, 8D11 identified a small, discrete population of CD45+ cells within the peripheral blood, cord blood, and BM of sheep. This population comprised 0.5–2.5% of the total sheep BM MNC, a proportion in close accord with the previously reported incidence of CD34+ cells in adult human BM. The ability of 8D11 to enrich for sheep hematopoietic progenitors was tested by magnetically sorting 8D11+cells and assessing colony-forming potential (CFU) and the CAFC frequency. Ovine CD34+ cells were 35- to 150-fold enriched for CFU and CAFC as compared with BM mononuclear cells, whereas CD34-negative cells were correspondingly depleted of progenitors. G-CSF mobilization of sheep resulted in a 14-fold increase in the levels of circulating CD34+ cells on day 4, providing further evidence of the utility of 8D11 as a marker of primitive hematopoietic cells in the sheep model. Antibody 8D11 also robustly labeled the lining of blood vessels in both frozen and paraffin-embedded sheep tissues, further extending the utility of this antibody to include analysis of tissue sections. In conclusion, this first successful generation of a monoclonal antibody to sheep CD34 will greatly facilitate the use of the sheep as a clinically relevant large animal model system to study allogeneic HSC transplantation both in utero and in post-natal recipients using bone marrow, mobilized peripheral blood, and cord blood as cell sources.

scholarly journals Distinct cellular immune profiles in the airways and blood of critically ill patients with COVID-19

Thorax ◽  
2021 ◽  
pp. thoraxjnl-2020-216256
Author(s):  
Anno Saris ◽  
Tom DY Reijnders ◽  
Esther J Nossent ◽  
Alex R Schuurman ◽  
Jan Verhoeff ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cells ◽  
T Cell ◽  
T Cell Activation ◽  
Inflammatory Mediators ◽  
Peripheral Blood ◽  
Cell Activation ◽  
Mononuclear Cells ◽  
Effector Memory ◽  
Activated T Cells

BackgroundKnowledge of the pathophysiology of COVID-19 is almost exclusively derived from studies that examined the immune response in blood. We here aimed to analyse the pulmonary immune response during severe COVID-19 and to compare this with blood responses.MethodsThis was an observational study in patients with COVID-19 admitted to the intensive care unit (ICU). Mononuclear cells were purified from bronchoalveolar lavage fluid (BALF) and blood, and analysed by spectral flow cytometry; inflammatory mediators were measured in BALF and plasma.FindingsPaired blood and BALF samples were obtained from 17 patients, four of whom died in the ICU. Macrophages and T cells were the most abundant cells in BALF, with a high percentage of T cells expressing the ƴδ T cell receptor. In the lungs, both CD4 and CD8 T cells were predominantly effector memory cells (87·3% and 83·8%, respectively), and these cells expressed higher levels of the exhaustion marker programmad death-1 than in peripheral blood. Prolonged ICU stay (>14 days) was associated with a reduced proportion of activated T cells in peripheral blood and even more so in BALF. T cell activation in blood, but not in BALF, was higher in fatal COVID-19 cases. Increased levels of inflammatory mediators were more pronounced in BALF than in plasma.InterpretationThe bronchoalveolar immune response in COVID-19 has a unique local profile that strongly differs from the immune profile in peripheral blood. Fully elucidating COVID-19 pathophysiology will require investigation of the pulmonary immune response.

scholarly journals Radiosensitivity of peripheral blood B cells in pigs

2009 ◽  
Vol 54 (No. 5) ◽  
pp. 223-235 ◽  
Author(s):  
Z. Sinkorova ◽  
J. Sinkora ◽  
L. Zarybnicka ◽  
Z. Vilasova ◽  
J. Pejchal
Keyword(s):  
B Cells ◽  
B Cell ◽  
Peripheral Blood ◽  
Dose Range ◽  
Whole Body ◽  
Large Animal Model ◽  
Large Animal ◽  
Cell Subpopulations

: Swine are here introduced to biodosimetry in an attempt to develop a large animal model allowing for comparison of <I>in vitro</I> experiments with the <I>in vivo</I> processes occurring after exposure to gamma radiation. This work investigates the radiosensitivity of the B cell compartment in peripheral blood. Four-week-old piglets were irradiated using the whole body protocol or full blood samples were irradiated <I>in vitro</I> in the dose range of 0–10 Gy. Relative radioresistance of B cell subpopulations and subsets was determined by measuring their relative numbers in leukocyte preparations at selected time intervals after irradiation using two color immunophenotyping and flow cytometry. Porcine B cells represent the most radiosensitive lymphocyte population in peripheral blood. Among B cell subpopulations and subsets investigated, the CD21+SWC7+ and CD21+CD1+ cells are highly radiosensitive and possess biodosimetric potential, at least in the range of low doses. Differences between cultures irradiated <I>in vitro</I> and lymphocyte dynamics in peripheral blood of irradiated animals clearly document the limits of <I>in vitro</I> data extrapolation in biodosimetry. We have shown that pigs can successfully be used in radiobiology and experimental biodosimetry due mainly to their availability, size and a relatively broad spectrum of available immunoreagents for lymphocyte classification.

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