• Korean Journal of Veterinary Research
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• v.29 no.4
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• pp.525-530
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• 1989

In order to enumerate the T-lymphocytes in bovine peripheral blood lymphocytes (PBL) by E rosette assay, KGRBC were treated with various concentrations of 2-aminoethylisothiouronium bromide(AET) and dextran(Dex), singly or in combination. To further standardize the assay, optimum concentration of AET- and/or Dex-treatment and incubation time for rosette forming cell(RFC) counts were determined. The levels of B-lymphocytes in the PBL were evaluated by erythzocyte-antibody($EA_{Fc}$)- and erythrocyte-antibody-complement (EAC)-rosetting techniques. The results obtained were as follows; The PBL from 20 clinically normal Korean cattle were formed as low percentage of spontaneous E-rosette ($6.7{\pm}2.4%$) in control group, whereas in KGRBC treated with 0.1M AET for 20 minutes and 8% Dex were formed as $37.3{\pm}2.7%$ and $45.1{\pm}2.1%$, respectively. And the synergistic effects were noted no less than $66.5{\pm}5.6%$ when the KGRBC treated with 0.1M AET and 8% Dex subsequently and rate of RFR did not change significantly between 3~24 hours incubation time at $4^{\circ}C$, EA-and EAC-RFR were $23.3{\pm}9.1%$ and $23.1{\pm}7.9%$, respectively. These results suggest that the KGRBC would be a useful agent for the enumeration of T-lymphocytes by E rosette assay and B-lymphocytes by EA- or EAC-rosette assay in cattle-PBL.

• Korean Journal of Veterinary Research
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• v.27 no.2
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• pp.253-258
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• 1987

The erythrocytes(E) rosette forming capacity of peripheral blood lymphocytes(PBL) in Korean native cattle was determined with 2-aminoethylisothiouronium bromide hydrobromide(AET) and dextran(Dex)-treated sheep erythrocytes(SRBC). To further standardize the assay, optimum concentration of AET and/or Dex-treatment and incubation time for rosette forming cell(RFC) counts were determined. In untreated SRBC resuspended in the rosetting medium(RPMI 1640 containing 10% FCS), PBL from 7 animals formed low percentage of rosettes($7.8{\pm}6.0%$). Both AET and Dex treatment not only enhanced the rosette formation but also made it easy to enumerate rosettes by increasing numbers of SRBC attached on them. SRBC treated with 0.1M AET for 20 min or 8% Dex formed the highest percentages of rosettes, ($35.7{\pm}6.0%$ and $48.3{\pm}4.7%$, respectively) and were used in subsequent studies. With SRBC treated with 0.1M AET for 20 min and suspended in 8% Dex, the maximum RFC observed were $56.6{\pm}6.8%$ in female and $65.8{\pm}6.3%$ in male and the rates of RFC did not change significantly between 3 and 20 hr incubation time at ${4^{\circ}C}$.