RESUMO
We report the use of a new surgical technique in a male dog affected by extensive stenosis of intrapelvic urethra through a antepubic urethral deviation, as an alternative to prepubic urethrostomy and ablation of the external genitalia. The technique consisted initially of orchiectomy, followed by retroumbilical celiotomy, transverse section of the penis in the pre-scrotal region and transposition of this towards the abdominal cavity by making anastomosis to the prostatic urethra. The dog was evaluated clinically and by urethrography retrograde positive contrast for up to four years after the procedure without any clinical signs, changes in urine stream or stenosis image. It is concluded that the pre-pubic urethral transposition is a viable alternative treatment for this dog affected by extensive stenosis of the membranous urethra.(AU)
Assuntos
Animais , Masculino , Cães , Anastomose Cirúrgica/veterinária , Estreitamento Uretral/cirurgia , Estreitamento Uretral/veterinária , Uretra/cirurgiaRESUMO
We report the use of a new surgical technique in a male dog affected by extensive stenosis of intrapelvic urethra through a antepubic urethral deviation, as an alternative to prepubic urethrostomy and ablation of the external genitalia. The technique consisted initially of orchiectomy, followed by retroumbilical celiotomy, transverse section of the penis in the pre-scrotal region and transposition of this towards the abdominal cavity by making anastomosis to the prostatic urethra. The dog was evaluated clinically and by urethrography retrograde positive contrast for up to four years after the procedure without any clinical signs, changes in urine stream or stenosis image. It is concluded that the pre-pubic urethral transposition is a viable alternative treatment for this dog affected by extensive stenosis of the membranous urethra.(AU)
Assuntos
Animais , Masculino , Cães , Estreitamento Uretral/veterinária , Anastomose Cirúrgica/veterinária , Estreitamento Uretral/cirurgia , Uretra/cirurgiaRESUMO
A palmitate-conjugate derivative of ovalbumin which can be inserted into the membrane of B cells has been prepared. The ability of these cells to act as antigen-presenting cells for specific T lymphocytes obtained from immunized mice was tested. It was found that the conjugates were more efficiently processed and presented than the naive form of the antigen. Palmitate-conjugated antibodies specific to ovalbumin were also inserted into the cell membrane of normal B lymphocytes. These cells were pulsed with the antigen and tested as antigen-presenting cells for T cells obtained from immunized mice. The antibody-decorated B cells presented ovalbumin more efficiently than non-decorated controls. Whether antibody-decorated, antigen-pulsed B cells could prime T cells in vivo was investigated. Some priming activity was found.
Assuntos
Apresentação de Antígeno/imunologia , Linfócitos B/imunologia , Receptores de Superfície Celular/imunologia , Linfócitos T/imunologia , Animais , Células Apresentadoras de Antígenos/imunologia , Células Cultivadas , Feminino , Imunização , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
Palmitate-conjugated monoclonal antibodies specific to ovalbumin were inserted into the cell membrane of normal resting B cells and LPS-activated blasts. These two decorated B cells were tested for their ability to act as antigen-presenting cells for ovalbumin-specific I-Ad-restricted T-cell hybridomas. It was found that the antibody-decorated resting B cells presented antigen more efficiently than non-decorated controls. However, no increment was observed when decorated LPS blasts were compared with non-decorated blasts. This is explained by the fact that the inserted antibodies quickly disappeared from the cell membrane of LPS blasts, while they were retained for a long period in the membrane of resting B cells.
Assuntos
Células Apresentadoras de Antígenos/fisiologia , Linfócitos B/fisiologia , Lipopolissacarídeos , Ativação Linfocitária , Receptores Imunológicos/fisiologia , Animais , Células Cultivadas , Feminino , Masculino , Camundongos , Camundongos Endogâmicos BALB CRESUMO
We have introduced some modifications in the technique called "cell decoration" in order to increase the amount of lipid-conjugated antibodies which can be incorporated into the membrane of B cells. As shown by FACS analysis, we have obtained an approximately 4-fold increment in the amount of specific antibodies incorporated into the cell membrane. The procedure, which consists of successive changes of the medium that contains the lipid-conjugated antibodies, avoided changes on parameters that interfere with cell viability. The proposed modification resulted in an approximately 2-fold enhancement of the ability of decorated B cells to act as antigen presenting cells for specific T hybridomas.
Assuntos
Anticorpos/imunologia , Células Apresentadoras de Antígenos/imunologia , Linfócitos B/imunologia , Lipídeos de Membrana/imunologia , Animais , Anticorpos/efeitos dos fármacos , Especificidade de Anticorpos/efeitos dos fármacos , Especificidade de Anticorpos/imunologia , Células Apresentadoras de Antígenos/efeitos dos fármacos , Linfócitos B/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Membrana Celular/imunologia , Ácido Desoxicólico , Técnicas Imunológicas , Indicadores e Reagentes , Camundongos , Camundongos Endogâmicos BALB C , Receptores de Antígenos/efeitos dos fármacos , Receptores de Antígenos/imunologia , SolubilidadeRESUMO
We have introduced some modifications in the technique called "cell decoration" in order to increase the amount of lipid-conjugated antibodies which can be incorporated into the membrane of B cells. As shown by FACS anlysis, we have obtained an approximately 4-fold increment in the amount of specific antibodies incroporated into the cell membrane. The procedure, which consists of successive changes of the medium that contains the lipid-conjugated antibodies, avoided changes on parameters that interfere with cell viability. The proposed modification resulted in an approximately 2-fold enhancement of the ability of decorated B cells to act as antigen presenting cells for specific T hybridomas
Assuntos
Anticorpos , Antígenos , Linfócitos B , Membrana Celular , Lipídeos , HibridomasRESUMO
Monoclonal antibodies specific for ovalbumin were conjugated to palmitate and inserted into the membrane of normal spleen B cells. Their presence in the membrane, as well as their ability to bind ovalbumin, was established by immunofluorescence. The so called anti-ovalbumin-'decorated' B cells were tested for their ability to act as antigen-presenting cells for ovalbumin-specific I-Ad-restricted T-cell hybridomas. It was found that the antibody-decorated B cells presented antigen more efficiently than non-decorated B cells.
Assuntos
Células Apresentadoras de Antígenos/imunologia , Linfócitos B/imunologia , Receptores de Antígenos de Linfócitos B/fisiologia , Receptores Imunológicos/fisiologia , Animais , Feminino , Hibridomas/imunologia , Imunoglobulina M/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Ovalbumina/imunologiaRESUMO
We have investigated the ability of different cells from non-immunized mice of the BALB/c strain to present antigen to two ovalbumin-specific I-Ad-restricted T hybridomas. Lipopolysaccharide-activated B-cell blasts were found to be the most efficient antigen-presenting cells. Purified small and dense splenic B cells also stimulated the hybridomas, although not to the same extent as the activated blasts, but comparable to non-fractionated spleen cells. Glutaraldehyde-treated B cells failed to present antigen, whereas F(ab')2 anti-mouse IgM-treated B cells exhibited markedly increased ability to present antigen. Using flow cytometry, we further purified the resting B cells by sorting the small lymphocytes to ensure that the ability of these cells to activate the hybridomas was not due to contamination with large non-resting B cells. The sorted small B cells retained the ability of antigen presentation. Their resting state was confirmed by the fact that they did not incorporate [3H]-thymidine as shown by autoradiographic analysis.