Blood typing in positive DEA 1 dogs: comparative analysis between immunochromatography, hemagglutination and flow cytometry / Tipificação sanguínea em cães AEC 1 positivo: análise comparativa entre a imunocromatografia, hemoaglutinação e citometria de fluxo

Blood typing techniques have been improved to ensure greater safety for transfusion procedures. Typification for the DEA 1 antigen through flow cytometry should offer more reliability to routine immunohematology in donor and recipient dogs. Currently, the DEA 1 group is starting to be an autosomal dominant allelic system with the DEA 1 negative type and its variations of positivity. The present study investigated the DEA 1 antigen using the techniques of immunochromatography, hemagglutination and flow cytometry. Among the positive animals for the DEA 1 group, typified by flow cytometry, medium intensities of fluorescence were found, which are indicative of weak, moderate and strong antigenicity. This enabled the division of the DEA 1 group into weak positive, moderate positive and strong positive. The blood typing techniques for the DEA 1 group by flow cytometry, agglutination and immunochromatography had positive (Spearman r=0.70) and statistically significant (p>0.0001) correlations.(AU)

As técnicas de tipificação sanguínea vêm sendo aperfeiçoadas para garantir maior segurança aos procedimentos transfusionais. A tipificação para o antígeno AEC 1 com o emprego da citometria de fluxo poderá oferecer mais confiabilidade à rotina da imunohematologia em cães doadores e receptores. Na atualidade, o grupo AEC 1 passou a ser denominado como um sistema alélico autossômico dominante com o tipo AEC 1 negativo e suas variações de positividade. O presente trabalho comparou os resultados de três técnicas utilizadas para a pesquisa do antígeno AEC 1: cromatografia; hemoaglutinação e citometria de fluxo. Dentro dos indivíduos positivos para o grupo AEC 1, tipificados pela citometria de fluxo, foram encontradas intensidades médias de fluorescência indicadoras de antigenicidade fraca, moderada e forte, podendo-se dividir o grupo AEC 1 em positivo fraco, positivo moderado e positivo forte. As técnicas de tipificação sanguínea para o grupo AEC 1 por cromatografia, hemoaglutinação e citometria de fluxo apresentaram correlação positiva (Spearman r=0,70) e estatisticamente significativa (p<0,0001).(AU)

Frequency of dog erythrocyte antigen 1 blood group and risk of incompatible transfusion in dogs of different breeds and mongrels from the city of Salvador - BA, Brazil / Frequência do antígeno eritrocitário canino 1 e risco de transfusão incompatível em cães de diferentes raças e mestiços da cidade de Salvador-BA, Brasil

The dog erythrocyte antigen 1 (DEA 1) is the most immunogenic blood group in dogs, and blood transfusions may trigger some undesirable effects in veterinary patients, which are directly associated with incompatible transfusions. The present study aimed to investigate the frequency of positive DEA 1 blood group in blood donor dogs from a blood bank in Salvador, Bahia, Brazil, and also to calculate the risk of managing incompatible blood in both first and second transfusion. A number of 203 dogs of different breeds, aged between 1 and 8 years, weighing 28 kg, with no degree of kinship and of both sexes in Salvador - BA, Brazil were evaluated to investigate the blood type DEA 1 frequency, by means of chromatography and flow cytometry tests for blood typing. The risk of incompatible blood transfusion in either a first or a second transfusion was also calculated. The frequency of the DEA 1 group ranged from 0% to 100% in various breeds, but with a mean positivity of 62.07% (126/203). And the lowest risk of an DEA 1 negative animal receiving DEA 1 positive blood within the group of animals evaluated was 0.92% at a first transfusion; and the risk of the same animal receiving incompatible blood for the DEA group 1 in the second transfusion was 0.008%. The highest risk of an DEA 1 negative animal receiving DEA 1 positive blood from these animals was 69.12%; and the risk of receiving incompatible blood for DEA 1 was 47.77%. In conclusion, the frequency of the DEA 1 group varied between the studied breeds and the risk of incompatible blood transfusions varies according to donor and recipiente breeds, but this can be overridden if blood typing tests are performed along with the cross-reaction test for compatibility.

O antígeno eritrocitário canino 1 (AEC 1) é o grupo sanguíneo mais imunogênico em cães, podendo as transfusões sanguíneas desencadearem alguns efeitos indesejáveis nos pacientes veterinários. Estes estão diretamente associados à transfusões incompatíveis. O presente trabalho teve como objetivo estudar a frequência do grupo sanguíneo AEC 1 em cães doadores de sangue de um banco de sangue de Salvador-BA, Brasil, e calcular o risco de administrar sangue incompatível tanto em uma primeira quanto em uma segunda transfusão. Foram avaliados 203 cães de diversas raças, de ambos os sexos, com idade entre 1 e 8 anos, peso a partir de 28 kg, sem nenhum grau de parentesco originários de Salvador ­ BA, Brasil, para pesquisa da frequência do tipo sanguíneo AEC 1, por meio de testes de imunocromatografia e citometria de fluxo para tipagem sanguínea. E calculado o risco de transfusão sanguínea incompatível tanto em uma primeira como em uma segunda transfusão. A frequência do grupo AEC 1 variou entre as raças estudadas de 0% a 100%, porém com uma positividade média de 62,07% (126/203). O menor risco de um animal AEC 1 negativo receber sangue AEC 1 positivo, dentro do grupo dos animais avaliados foi de 0,92% em uma primeira transfusão e o risco do mesmo animal receber sangue incompatível para o gruo AEC 1 na segunda transfusão foi de 0,008%. Quanto ao maior risco de um animal AEC 1 negativo receber sangue AEC 1 positivo destes animais foi de 69,12% e o risco do mesmo receber sangue incompatível para o AEC 1 foi de 47,77%. A frequência do grupo AEC 1 variou entre as raças estudadas e o risco de transfusões incompatíveis variou de acordo com as raças doadoras e receptoras, mas esse risco pode ser anulado se sempre forem realizados os testes para tipagem sanguínea junto com a prova de reação cruzada para compatibilidade.

Detection of canine anti-DEA 1 antibodies using flow cytometry in dogs following DEA 1-positive blood transfusion / Detecção de anticorpos caninos anti-AEC 1 por citometria de fluxo em cães pós-transfundidos com sangue AEC 1 positivo

The shortage of dog blood donors in veterinary emergencies can lead to blood transfusions between animals whose blood type has not been identified. The antibody profile serves as a warning sign for animals that require a second blood transfusion, which is only advisable from compatible donor dogs. This article focuses on determination of anti-DEA 1 antibodies using the flow cytometry technique in dogs that have undergone a transfusion using DEA 1-positive blood, compared to results obtained from crossmatching. Blood from 18 DEA 1-positive donors ranked according to the chromatographic technique was used to transfuse thirty-three animals with unknown blood types and which demonstrated negative crossmatching to donors. On post-transfusion days 7, 14, 21 and 28, 45% and 27% of the animals tested positive for the anti-DEA 1 antibody, through crossmatching and flow cytometry, respectively. Detecting antibodies using the flow cytometric technique has high specificity and sensitivity, while crossmatching methods are highly sensitive but manifest low specificity. Following blood transfusion, animals that did not present as positive through crossmatching or flow cytometry were considered different from all other DEA 1-positive blood groups.(AU)

A escassez de cães doadores de sangue em situações de emergência na Medicina Veterinária pode levar a realização de transfusões de sangue entre animais que não tiveram seu tipo sanguíneo previamente determinado. O padrão de anticorpos serve como um sinal de alerta para animais que serão submetidos a uma segunda transfusão sanguínea, sendo essa somente recomendável a partir de cães doadores compatíveis. Este artigo aborda a pesquisa de anticorpos anti-AEC 1 pela técnica de citometria de fluxo em cães que receberam uma transfusão utilizando sangue do grupo AEC 1 positivo, comparando os resultados com aqueles obtidos a partir de reação cruzada. Foi utilizado sangue de 18 animais doadores do tipo AEC 1 positivo classificados por técnica cromatográfica para transfundir trinta e três animais com tipos sanguíneos desconhecidos, os quais mostraram reação cruzada negativa aos doadores. Nos dias 7, 14, 21 e 28 pós-transfusão, 45% e 27% dos animais mostraram-se positivos para os anticorpos anti-AEC 1, respectivamente, pela reação cruzada e através de citometria de fluxo. A pesquisa de anticorpos com o emprego da técnica de citometria de fluxo tem alta especificidade e sensibilidade, enquanto a reação cruzada, altamente sensível, tem baixa especificidade. Animais que não apresentaram positividade após a transfusão de sangue na reação cruzada e na citometria de fluxo concomitantemente foram considerados de qualquer outro grupo sanguíneo diferente do grupo sanguíneo AEC 1 positivo.(AU)

Evaluation of dry eye syndrom in C57BL/6.NOD-Aec1Aec2 mice / 中华实验眼科杂志

Background C57BL/6.NOD-Aec1Aec2 mouse is considered to be an idea model for the study of the pathogenesis of Sj(o)gren syndrome,but the cause of dry eye in these mice is unclear.Objective This study was to investigate the histopathological change of the ocular surfaces of C57BL/6.NOD-Aec1Aec2 mice,and to determine whether dry eye is developed spontaneously in these mice.Methods Forty-five clean C57BL/6.NOD-Aec1 Aec2 mice were used as the experiment group and forty-five C57BL/6J mice(both male and female)were used as the control group in this study.Detection of fasting blood-glucose,Schirmer' s test Ⅰ (S Ⅰ t),lissamine green staining and scoring of the corneal and conjunctival epithelium in the mice were performed at the age of 4,8,12,16 and 20weeks.Five mice from each group were sacrificed and their corneas were obtained to measure the central corneal epithelium thickness and to count the number of conjunctival goblet cells.In addition,lymphocyte infiltration in the lacrimal gland of the mice was examined with hematoxylin-eosin staining.The uhrastructure of the corneal epithelial cells and microvilli were assessed by scanning electron microscopy.The use and care of the mice were approved by the Experimental Animal Care Committee of the Third Military Medical University.Results No sign of dry eye was seen in both the 4-week-old C57BL/6.NOD-Aec1Aec2 mice and 4-week-old C57BL/6J mice.The S Ⅰ t values in 8-week-old,12-week-old,16-week-old and 20 week-old mice from the experiment group were (2.7 ±0.9) mm,(2.5 ±0.8) mm,(1.8±0.6) mm and (1.9± 0.1) mm,respectively,showing a significant reduction in comparison with those of the control mice of the same age(all P＜0.01).The amount of lissamine green staining in the C57BL/6.NOD-Aec1Aec2 mice gradually increased with age,showing elevated scores in 12-week-old,16-week-old and 20-week-old mice in the experiment group(all P＜0.01).The central corneal epithelium thicknesses were(20.18±3.75)μm,(17.01 ±5.25) μm,(14.19±5.72) μm and(12.00±3.25) μm in the 8-week-old,12-week-old,16-week-old and 20-week-old C57BL/6.NOD-Aec1Aec2 mice,respectively,which were significantly lower than those of the C57BL/6Jmice of the same age (all P＜0.01).The numbers of conjunctival goblet cells were (8.2±2.4),(6.2±2.1),(6.1 ±2.2) and (4.1 ± 2.0) in the 8-week-old,12-week-old,16-week-old and 20-week-old C57 BL/6.NOD-Aec 1Aec2 mice,respectively,showing a gradual decrease with age and a significant decline in comparison with those of the C57BL/6Jmice of the same age(all P＜0.01).Lymphocyte infiltration in the lacrimal gland and destruction of gland ducts were seen by hematoxylin-eosin staining,and acinar abnormality aggravated with aging.Reduction of corneal epithelial cells and the number of microvilli were distinguished with aging under the scanning electron microscope.The fasting bloodglucose levels of the two groups were both less than 6.0 mmol/L,and no significant difference was found between them at any age(P=0.637,0.610,0.163,0.086,0.938).Conclusions C57BL/6.NOD-Aec1Aec2 mice develop dry eye spontaneously with aging.The course of disease and characteristics of dry eye in C57BL/6.NOD-Aec1Aec2mice is similar to human dry eye.The C57BL/6NOD-Aec1 Aec2 mouse is the perfect model to study the pathogenesis of dry eye.