This study is the first to provide information on the lymphocyte subpopulations in peritoneal effusions in horses. Peritoneal transudates (n = 12), peritoneal exudates (n = 6) and a pleural exudate (n = 1) were analyzed. The total nucleated cell count (TNCC), total protein (TP) and matrix metalloproteinase-9 (MMP-9) concentration determined by ELISA were measured and routine cytological evaluation was performed. CD3, CD4, CD8 and CD21 positive cells were detected by flow cytometry. A higher percentage of neutrophils (P < 0.05) and higher MMP-9 (P < 0.01) levels were found in exudates. A higher percentage of macrophages (P < 0.05) and lymphocytes (P < 0.01) were found in transudates. CD4 + lymphocytes were the most common lymphocyte subpopulation in all samples. CD21 + lymphocytes were the least common in all samples. A large variability in the percentage of CD21 + lymphocytes was found in exudates. The percentage of CD21 + lymphocytes positively correlated with the level of total protein (r = 0.5704, P < 0.05). The correlation was even stronger in the group of exudates. The percentages of lymphocyte subpopulations did not correlate with the level of MMP-9 or with cytological findings. The level of MMP-9 positively correlated with the percentage of neutrophils (r = 0.4980, P < 0.05), the level of TP (r = 0.7855, P < 0.01) and TNCC (r = 0.6129, P < 0.01). A significantly higher level of MMP-9 was detected in euthanized horses than in horses that survived (P < 0.05). However, it was shown that the level of MMP-9 in the peritoneal fluid can change significantly in a short time. More studies on repeated abdominocentesis could contribute to elucidating the role of MMP-9 as a prognostic indicator.
- Klíčová slova
- Abdominocentesis, B lymphocytes, Eosinophils, Peritoneal fluid, Survive, T lymphocytes, Tc lymphocytes, Th lymphocytes,
- MeSH
- CD4-pozitivní T-lymfocyty MeSH
- exsudáty a transsudáty * MeSH
- imunofenotypizace veterinární MeSH
- koně MeSH
- matrixová metaloproteinasa 9 * MeSH
- počet buněk veterinární MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- matrixová metaloproteinasa 9 * MeSH
Equine sarcoids (ES) are known globally as the most frequent skin tumour affecting horses. These tumours affect the horse's monetary value, they can affect the horse's welfare and can be difficult and expensive to treat. Bovine papillomavirus (BPV) is considered to be the aetiological agent of this tumour, as BPV 1, 2 and 13 have been detected in ES. This is the only known natural cross species infection by a papillomavirus. The BPV genome can be divided into two coding regions: The early region E which encodes the transforming proteins E5, E6 and E7 as well as the replication and transcription regulatory proteins E1 and E2 and the late region encoding the structural proteins of the virus L1 and L2. The E5 oncoprotein is believed to downregulate MHC 1 and as a result, escapes an immune reaction with affecting the cells cycle and eventually allows the viral affected cells to proliferate into ES. We have constructed an ELISA test by utilising the C terminal peptide of the E5 oncoprotein and explored the possibility of IgG antibodies existence in horses to the E5 oncoprotein. For this study we have examined 136 horses, some showing ES lesions (80 horses) and some without ES lesions (56 horses). By using our ELISA test, we have shown that antibodies to the E5 oncoprotein are in fact present and that from a certain level seem to be found only in ES positive horses. Therefore, proving that an immune response to this protein can be expected.
- Klíčová slova
- Bovine papillomavirus, E5 oncoprotein, ELISA, Equine sarcoid, MHC,
- Publikační typ
- časopisecké články MeSH
Lymphocyte immunophenotyping can be useful for evaluating immune competence and predicting the disease prognosis. It is essential to gain knowledge about canine lymphocyte immunophenotypes in various conditions. The study deals with the characteristics of lymphopenia in dogs, with an emphasis on lymphocyte immunophenotyping by flow cytometry. Blood samples from 44 dogs with lymphopenia were included in the study. All lymphopenias sent from veterinary clinics to the diagnostic laboratory were analyzed. The hematological and biochemical abnormalities were investigated, as well as the effect of the age. Lymphopenias were classified according to the level of C-reactive protein (CRP). The percentage of T cells, B cells, Th cells and Tc cells, and T/B and Th/Tc ratios were determined by flow cytometry. Lymphopenias often occurred in dogs over 7 years of age (79.5 %). The most common were postoperative lymphopenia (31.8 %) and inflammatory diseases (29.5 %), most commonly affecting the gastrointestinal tract. Frequent abnormalities were monocytosis (56.8 %), increased CRP (72.7 %) and decreased albumin/globulin ratio (50.0 %). The percentage of Th lymphocytes was significantly lower in the group with elevated CRP than in the group with basal CRP (P = 0.0329). A negative correlation was found between the level of CRP and the percentage of Th lymphocytes (r = -0.3278, P = 0.0390). This study provided new insights into the appearance, incidence and classification of canine lymphopenia.
- Klíčová slova
- Acute inflammation, Aging dogs, C-reactive protein, Flow cytometry, Stress,
- MeSH
- B-lymfocyty MeSH
- imunofenotypizace veterinární MeSH
- lymfocyty MeSH
- lymfopenie * diagnóza veterinární MeSH
- nemoci psů * diagnóza MeSH
- průtoková cytometrie veterinární MeSH
- psi MeSH
- T-lymfocyty MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Meloxicam is a commonly used analgesic in rabbits. However, its possible impact on lymphocyte subpopulations remained unknown. The aim of the study was to investigate a possible effect of long-term administration of meloxicam on rabbit lymphocyte subpopulations. The study included 8 rabbits given meloxicam orally once daily (1 mg/kg BW) for 14 days and 8 rabbits as a control group. Peripheral blood samples were collected on day 0 (before the first dose of meloxicam), day 3, 7 and 14. Samples were evaluated with a haematology analyser and a flow cytometer. A significant decrease in T: B cell ratio was found in all samples taken during meloxicam administration compared to day 0, as well as in comparison with the control group (P < 0.01). A significant increase (P < 0.05) in proportion of CD5 +CD8 + lymphocytes occurred by day 3. Subsequently, although the values slightly decreased, they still remained elevated throughout all the experiment compared to the values from day 0 (P < 0.05). A slight decrease in T and B cell activation (CD5 +CD25 + and IgM+CD25 +) noticed by day 3, declined during the next days of administration and became more and more significant (finally, P = 0.0078). Since a high significant decrease (P < 0.01) in both T and B cell activation as well as a significant increase (P < 0.05) in CD5 +CD8 + T cells proportion were observed after meloxicam administration, a predicted effect of long-term administration of meloxicam on rabbit lymphocytes was confirmed.
- Klíčová slova
- CD25, CD8 T cells, Flow Cytometry, NSAIDs, T: B cell ratio,
- MeSH
- aktivace lymfocytů MeSH
- CD8-pozitivní T-lymfocyty * MeSH
- králíci MeSH
- meloxikam farmakologie MeSH
- podskupiny lymfocytů * MeSH
- průtoková cytometrie veterinární MeSH
- zvířata MeSH
- Check Tag
- králíci MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- meloxikam MeSH
Ferrets are nowadays frequently used as animal models for biomedical purposes; in many cases, immunosuppression of experimental animals is necessary. The aim of this study was to evaluate the effect of intramuscular dexamethasone administration (2 mg/kg as the initiation dose continued with 1 mg/kg q 12 h applied 5 times) on ferret's immune system. In comparison with ferrets which received the saline (n = 5), significantly lower total counts of leukocytes (P < 0.01), lymphocytes (P < 0.01) and monocyte (P < 0.05), as well as absolute numbers of CD4+CD8- (P < 0.01) and CD4-CD8+ (P < 0.01) subsets were noted in dexamethasone treated ferrets (n = 5) the first day after the treatment (D1). Absolute number of CD79+ lymphocytes remained unchanged throughout the experiment. The proliferation activity of lymphocytes in dexamethasone treated ferrets was lower only in D1 using concanavalin A (conA), phytohemagglutinin (PHA) and pokeweed mitogen (PWM); statistical significance was noted using PHA 40 (P < 0.05) and PWM 10 (P < 0.01). Lower neutrophil activity (P < 0.01) was detected in D1 after the dexamethasone treatment in both production of reactive oxygen species (chemiluminescence test) and ingestion of particles (phagocytosis assay). The dexamethasone treatment proved to be useful for short-term immunosuppression in ferrets. The results closely resembled data previously reported in human studies and indicate classification of ferrets as steroid-resistant species.
- Klíčová slova
- Corticosteroids, Flow cytometry, Immunology, Leukocytes, Mustelidae,
- MeSH
- aktivace lymfocytů MeSH
- dexamethason * farmakologie MeSH
- fretky * MeSH
- fytohemaglutininy MeSH
- imunosupresivní léčba * veterinární MeSH
- mitogeny líčidla amerického MeSH
- modely u zvířat MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- dexamethason * MeSH
- fytohemaglutininy MeSH
- mitogeny líčidla amerického MeSH
The bovine genes TLR1, TLR2 and TLR6, which encode Toll-like receptors, key components of the innate immune system, were screened for polymorphisms in Czech Red Pied (Czech Simmental) cattle, and the different variants present in the population were tested for association with reproductive and fitness traits. Diversity was investigated in a group of 164 bulls using hybrid resequencing of pooled amplicons with PacBio technology and of pooled genomic DNA using HiSeq X-Ten technology. The validated single nucleotide polymorphisms (SNPs) were genotyped in individual animals using the primer extension technique. The association of genotypic classes of 16 polymorphisms with six phenotypic traits were estimated with one-way analysis of variance (ANOVA) and with restricted maximum likelihood (REML) algorithm. The evaluated traits included the incidence of cystic ovaries, index of early reproductive disorders, paternal and maternal indicators of calving ease, production longevity and calf vitality index. The estimated breeding values were used for combined trait quantification. Early traits, namely, cystic ovaries and early reproductive disorders, were not associated with any of the tested polymorphisms according to the general ANOVA test. By contrast, five variants of all three genes were associated with calving ease, both paternal and maternal. The production longevity correlated with two variants of TLR1 and the calf vitality index correlated with the 1044 T > C (rs68268249) polymorphism in TLR2. The false discovery rate (FDR) according to Benjamini-Hochberg was favourable for the calving ease trait (0.221) and maternal calving ease (0.214), which allows to consider the observed associations real, regardless of the error arising from the multiple comparisons. These results were supported by REML only partially, probably in view of the additivity assumption. Two mechanisms of action on calving are conceivable, either via infection resistance or via the involvement of TLR2 in signalling in the myometrium. The known formation of heterodimers by the TLR1, -2 and -6 products might be responsible for the shared pattern of action in these genes. The association of the calf vitality index with TLR2 variation might reflect the increased role of infections in calves compared to adult animals.
- Klíčová slova
- Cattle, Diversity, Effect prediction, Health traits, Toll-like receptors,
- MeSH
- chov MeSH
- dlouhověkost genetika MeSH
- fenotyp MeSH
- genotyp MeSH
- jednonukleotidový polymorfismus * MeSH
- rozmnožování genetika MeSH
- skot genetika MeSH
- toll-like receptor 1 genetika MeSH
- toll-like receptor 2 genetika MeSH
- toll-like receptor 6 genetika MeSH
- toll-like receptory genetika MeSH
- věkové faktory MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- skot genetika MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
- Názvy látek
- toll-like receptor 1 MeSH
- toll-like receptor 2 MeSH
- toll-like receptor 6 MeSH
- toll-like receptory MeSH
Several molecular clonality assays have been developed to assess canine B cell proliferations. These assays were based on different sequence data, utilized different assay designs and employed different testing strategies. This has resulted in a complex body of literature and complicates evidence-based selection of primer sets. In addition, further refinement of primer sets is difficult because it is unknown how well current primer sets cover the expressed sequence repertoire. The objectives of this study were 1) to provide an overview of published IGH clonality assays that highlights key differences in assay design and testing strategy and 2) to propose a novel method for optimizing primer sets that leverages large-scale sequencing data. A review of previously published assays highlighted confounding factors that hamper a direct comparison of performance metrics between studies. These findings illustrate the need for a multi-institutional effort to harmonize veterinary clonality testing. A novel in silico analysis of primer sequences using a large dataset of expressed sequences identified shortfalls of existing primer sets and was used to guide primer optimization. Three optimized primer sets were tested and yielded qualitative sensitivity values between 80-90%. The qualitative sensitivity ranged from 1% to over 50% and was dependent on the size of the neoplastic clone and the sample DNA used. These findings illustrate that inclusion of high-throughput sequencing data for primer design can be a useful tool to guide primer design and optimization. This strategy could be applied to other antigen receptor loci or species to further improve veterinary clonality assays.
- Klíčová slova
- B cell lymphoma, Clonality testing, Dog, Lymphoma, PARR, Sensitivity, Specificity,
- MeSH
- B-lymfocyty cytologie MeSH
- buněčné klony * MeSH
- DNA primery * MeSH
- psi genetika imunologie MeSH
- těžké řetězce imunoglobulinů genetika MeSH
- zvířata MeSH
- Check Tag
- psi genetika imunologie MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
- Názvy látek
- DNA primery * MeSH
- těžké řetězce imunoglobulinů MeSH
Events occurring in the chicken caecum following Salmonella Enteritidis infection are relatively well-described. However, mechanisms of the immune response and defence beyond the intestinal tract are less well-described. In this study, we therefore determined changes in protein abundance in the liver and blood serum in response to S. Enteritidis infection using the unbiased approach of shotgun proteomics. Complement and coagulation cascades, TNF signalling, antigen processing and presentation was activated in the liver following infection with S. Enteritidis. Chicken proteins that decreased in the liver were involved in glycolysis, the citrate cycle, oxidative phosphorylation and fatty acid metabolism. No functional category was significantly activated or suppressed in the serum. Concerning individual proteins, VNN1, SAA, AVD, SERPINA3, SERPINB10, AGT, MRP126 or CP increased in abundance both in the liver and serum. MT4, MT3, PTGDS, GLRX and TGM4, though highly inducible in the liver, did not increase in the serum. PIGR, SERPINF2 and IGJ increased in the serum but not in the liver. SERPINA4, apoAIV, CLEC3B, SERPINF1, HRG, AHSG and ALB decreased both in the liver and serum. Avidin-like LOC431660, THRSP, GATM, GGACT, ACOX1, ALDOB or FABP7 decreased in the liver but not in the serum. Finally, CKM, CKB, PLTP, COMP, IGFALS, AMY1A or SERPIND1 decreased in the serum after S. Enteritidis infection but not in the liver. Differently abundant proteins characterise the chicken's response to infection and can be also used as markers of chicken health status.
- Klíčová slova
- Acute phase protein, Chicken, Liver, Proteomics, Salmonella, Serum,
- MeSH
- cékum imunologie MeSH
- játra imunologie metabolismus mikrobiologie MeSH
- kur domácí krev imunologie MeSH
- nemoci drůbeže imunologie mikrobiologie MeSH
- prezentace antigenu MeSH
- proteiny akutní fáze analýza MeSH
- proteomika * MeSH
- Salmonella enteritidis MeSH
- salmonelová infekce u zvířat krev imunologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- proteiny akutní fáze MeSH
The ability to mount adaptive immune responses to a diverse array of pathogens is essential to maintaining the health of an individual. The outcome of adaptive immune responses is influenced by the pool of available lymphocyte antigen receptors. Understanding the composition and dynamics of immune repertoires is hence of relevance to characterizing physiologic immunological processes as well as understanding disease pathogenesis. The dog is increasingly recognized as a model for human disease. The objective of this study was to utilize NGS for comprehensive and unbiased analysis of the IGH repertoire in healthy dogs. First, the IGH locus was searched in silico for previously unidentified genes. Second, IGH transcripts from major lymphoid organs were amplified using a 5'RACE approach without V/J primer bias. Third, amplicons were sequenced on an Illumina MiSeq platform, and data were analyzed using the ARResT/Interrogate platform. Data analysis included V/J usage, V-J pairing biases, isotype frequency, CDR3 diversity, convergent recombination, and public repertoires. The results of this study provide a comprehensive IGH repertoire analysis for healthy dogs. These data will allow further improvement of V/J gene-specific primer sets and will serve as baseline for future studies investigating immune repertoires in health and disease.
- Klíčová slova
- Antigen receptor, CDR3, Dog, IGH, Rearrangement, Repertoire analysis,
- MeSH
- imunoglobulinové izotypy genetika MeSH
- psi MeSH
- receptory antigenů genetika MeSH
- sekvenční analýza DNA MeSH
- těžké řetězce imunoglobulinů genetika MeSH
- variabilní oblast imunoglobulinu genetika MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- zvířata MeSH
- Check Tag
- psi MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- imunoglobulinové izotypy MeSH
- receptory antigenů MeSH
- těžké řetězce imunoglobulinů MeSH
- variabilní oblast imunoglobulinu MeSH
Rheumatoid factor (RF) is used in human and veterinary medicine in the form of IgM RF traditionally to support the diagnosis of rheumatoid arthritis (RA). In the latest diagnostic criteria, presence of anti - citrullinated protein antibodies (ACPA) was added to the grading system for the diagnosis of RA in humans. A change which is not integrated or routinely used in veterinary medicine. The criteria changed partly because of RF's diagnostic shortcomings, which include its increased titer detection in humans with non-rheumatoid diseases, inability to predict the disease and increased titers over the limit in the older population. Clinical signs similar to human RA were reported in horses in a condition known as idiopathic polysynovitis. Similarities in the clinical presentation to RA raised a question to the usability of RF and ACPA in horses. In our study, sixty clinically healthy horses, ranging from 3 days to 30 years of age, were evaluated for their serum levels of IgM RF. 55 of these horses were tested for ACPA, using methods of ELISA measuring Anti - CFG (Anti citrullinated fibrinogen antibody). The results of the study demonstrated the existence of an age-dependent increase in the level of IgM RF up to the age of about 9 years and ACPA's independence of the horse's age as well as both markers independence of the horse sex.
- Klíčová slova
- ACPA, Age-dependency, ELISA, Horse, IgM RF, Polysynovitis,
- MeSH
- autoprotilátky krev MeSH
- biologické markery krev MeSH
- ELISA MeSH
- imunoglobulin G krev MeSH
- imunoglobulin M krev MeSH
- koně imunologie MeSH
- protilátky proti citrulinovaným peptidům krev MeSH
- revmatoidní artritida MeSH
- revmatoidní faktor krev MeSH
- synovitida diagnóza imunologie veterinární MeSH
- věkové faktory MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
- Názvy látek
- autoprotilátky MeSH
- biologické markery MeSH
- imunoglobulin G MeSH
- imunoglobulin M MeSH
- protilátky proti citrulinovaným peptidům MeSH
- revmatoidní faktor MeSH
