RESUMEN
Cell lines derived from spontaneous tumors serve as a research tool for cancer cell biology and new anti-cancer drug development. Isolation and propagation of canine lymphoma cell lines is difficult, thus only a few are available. Now we have established a new B-cell lymphoma cell line CLBL-1 from a dog with confirmed stage IV diffuse large cell lymphoma. Immunophenotyping of these CLBL-1 cells showed positive staining for CD11a, CD79alphacy, CD45, CD45RA, MHC II and cells were negative for CD3, CD4, CD5, CD8, CD11d, CD14, CD21, CD34, CD56 and T-cell receptor-gammadelta (TCR-gammadelta). PCR analysis for TCR-gamma and immunoglobulin heavy chain (IgH) gene rearrangements yielded a monoclonal result for the IgH gene. Furthermore, the clonality of IgH gene rearrangement was confirmed by sequencing of 16 positive bacterial clones. As canine lymphoma resembles non-Hodgkin's lymphoma (NHL) in humans in many respects, this new cell line, will promote translational and comparative lymphoma research in humans and dogs.
Asunto(s)
Linfocitos B/patología , Línea Celular Tumoral/patología , Enfermedades de los Perros/patología , Linfoma de Células B Grandes Difuso/veterinaria , Animales , Antígenos CD/análisis , Antígenos de Neoplasias/análisis , Linfocitos B/química , Biopsia con Aguja Fina , Línea Celular Tumoral/química , Separación Celular , Células Clonales/química , Células Clonales/patología , Perros , Citometría de Flujo , Reordenamiento Génico de Cadena Pesada de Linfocito B , Inmunofenotipificación , Ganglios Linfáticos/patología , Linfoma de Células B Grandes Difuso/patología , Masculino , Proteínas de Neoplasias/análisis , Receptores de Antígenos de Linfocitos T gamma-delta/análisisRESUMEN
The purpose of the study was to evaluate the effects on quantitative and qualitative microbial content of endoscopic bronchoalveolar lavage fluid (BALF) in healthy dogs using a laryngeal mask airway (LMA). It was hypothesised that oropharyngeal protection might prevent contamination of BALF with oropharyngeal microflora. Ten healthy Beagle dogs were randomly assigned to two groups and underwent endoscopic BAL on two occasions, either via an open unprotected oropharynx with oxygen supply provided via a nasal catheter (NT) or through a sterile LMA. For the second sampling, groups were switched. BALF analysis included quantitative microbial culture, nucleated cell counts and cytology. The mean (+/-SD) number of colony forming units (CFU)/mL found in the BALF using the LMA was 25,610+/-22,943 in the right lung (RL) and 22,510+/-18,779 in the left (LL). With the NT technique, the figures were 21,068+/-19,375 for the RL and 16,060+/-15,523 for the LL, respectively. Nucleated cell counts/microL were 691.0+/-181.6 (RL) and 734.0+/-171.6 (LL) for LMA, and 772.0+/-251.0 (RL) and 748+/-163.2 (LL) for NT. No significant differences were detected either in the number of CFU/mL or in the diversity of bacterial species with the two methods. A significant increase in BALF bacterial counts (with reduced species diversity) was observed on the second compared to the first sampling regardless of the method used. Protection of the oral cavity and oropharynx using an LMA had no significant influence on BALF bacterial counts. The findings suggest that with careful endoscope insertion, the risk of contamination of BALF by resident and transient oropharyngeal microflora can be negligible.
Asunto(s)
Bacterias Aerobias/aislamiento & purificación , Líquido del Lavado Bronquioalveolar/microbiología , Broncoscopía/veterinaria , Perros/microbiología , Manejo de Especímenes/veterinaria , Animales , Lavado Broncoalveolar/métodos , Lavado Broncoalveolar/veterinaria , Broncoscopía/métodos , Recuento de Colonia Microbiana/veterinaria , Femenino , Masculino , Oxígeno/sangre , Oxígeno/metabolismo , Distribución Aleatoria , Manejo de Especímenes/instrumentación , Manejo de Especímenes/métodosRESUMEN
The effects of pharmacological bronchoprovocation on airflow patterns and surrogate respiratory parameters assessed by barometric whole body plethysmography (BWBP) were investigated in healthy dogs, previously exposed to cadmium chloride inhalation. BWBP-derived respiratory variables were calculated (1) at baseline and (2) following nebulisation of increasing concentrations of histamine, carbachol and adenosine 5'-monophosphate (AMP) until enhanced pause (PENH) increased to 300% of baseline (PCPENH300). Bronchoalveolar lavage fluid (BALF) cytology before (BCC) and after (ACC) cadmium chloride inhalation revealed cadmium-induced airway inflammation. Neutrophils increased from 6.7 +/- 7.3% (728 +/- 104/microL) BCC to 77.8 +/- 8.6% (3255 +/- 1407/microL) ACC. PCPENH300 for all three agonists significantly decreased ACC (means+/-SD) as follows: PCPENH300(histamine) 0.72 +/- 0.28 mg/mL BCC, and 0.35 +/- 0.31 mg/mL ACC (P<0.02); PCPENH300(carbachol) 0.34 +/- 0.16 mg/mL BCC, and 0.064 +/- 0.032 mg/mL ACC (P<0.02); PCPENH300(AMP) 1000 mg/mL BCC, and 415 +/- 398 mg/mL ACC (P<0.03). The only clinical sign shown was coughing. It was concluded that airway hyperresponsiveness after induced airway inflammation can be determined by BWBP in conscious small sized dogs. BWBP could be a suitable technique to study the respiratory effects of urban environmental pollution in pets.