Petro T M, Schwartzbach S D, Zhang S
Department of Oral Biology, University of Nebraska Medical Center, Lincoln 68583-0740, USA.
Int J Immunopharmacol. 1999 Feb;21(2):103-14. doi: 10.1016/s0192-0561(98)00070-8.
To determine the influence of smokeless tobacco (ST) and nicotine on the cytokine phenotype of memory T-cells, splenic mononuclear cells (SPM) were exposed to 1:10(2) or 1:10(3) dilutions of ST extract (ST-SPM), 10 or 100 microg/ml nicotine (NIC-SPM), or medium (CON-SPM) during 4 days of stimulation with anti-CD3. SPM were then washed extensively to remove residual ST or nicotine and restimulated with anti-CD3 and anti-CD28 in the absence of ST or nicotine. Expression of IL-2, IL-4, IL-10 and IFN-gamma protein and mRNA levels after 4 days of primary stimulation and after 24 and 48 h of restimulation was evaluated using ELISA and RT-PCR, respectively. After 4 days of primary stimulation, SPM exposed to 100 microg/ml nicotine sustained expression of IL-2, IFN-gamma, IL-10 and IL-4 mRNA as opposed to CON-SPM. Restimulation of CON-SPM resulted in maximum re-expression of cytokine mRNA at 24 h and a decline by 48 h. Restimulated NIC-SPM in the absence of nicotine delayed maximal re-expression of IL-2, IFN-gamma, IL-10 and IL-4 mRNA until 48 h. Heightened expression of cytokine mRNA at 48 h was paralleled by a small but significant increase in production of IFN-gamma, IL-4 and IL-10 protein by NIC-SPM as measured by ELISA. In contrast, ST-SPM did not exhibit residual expression of cytokine mRNA after 4 days of primary stimulation. Like NIC-SPM, however, restimulated ST-SPM exhibited maximum IL-2, IL-4, IFN-gamma, and IL-10 mRNA at 48 h. Heightened re-expression of cytokine mRNA at 48 h by ST-SPM was paralleled by increased production of IL-2, IFN-gamma, IL-4 and IL-10 protein. These results indicate that exposure of T-cells to nicotine, but not ST, during a primary immune response result in inordinate cytokine expression after 4 days. In addition, memory T-cells initially exposed to nicotine or ST during a primary immune response, exhibit excessive cytokine expression when T-cells are restimulated in the absence of nicotine or ST. pharmacology.
为了确定无烟烟草（ST）和尼古丁对记忆T细胞细胞因子表型的影响，在抗CD3刺激的4天期间，将脾单核细胞（SPM）暴露于1:10²或1:10³稀释的ST提取物（ST-SPM）、10或100μg/ml尼古丁（NIC-SPM）或培养基（CON-SPM）中。然后将SPM充分洗涤以去除残留的ST或尼古丁，并在不存在ST或尼古丁的情况下用抗CD3和抗CD28再次刺激。分别使用ELISA和RT-PCR评估初次刺激4天后以及再次刺激24和48小时后IL-2、IL-4、IL-10和IFN-γ蛋白的表达以及mRNA水平。初次刺激4天后，与CON-SPM相比，暴露于100μg/ml尼古丁的SPM持续表达IL-2、IFN-γ、IL-10和IL-4 mRNA。CON-SPM的再次刺激导致细胞因子mRNA在24小时时最大程度地重新表达，到48小时时下降。在不存在尼古丁的情况下再次刺激的NIC-SPM将IL-2、IFN-γ、IL-10和IL-4 mRNA的最大重新表达延迟至48小时。通过ELISA测量，NIC-SPM在48小时时细胞因子mRNA的表达增强，同时IFN-γ、IL-4和IL-10蛋白的产生有小幅但显著的增加。相比之下，初次刺激4天后，ST-SPM未表现出细胞因子mRNA的残留表达。然而，与NIC-SPM一样，再次刺激的ST-SPM在48小时时表现出最大的IL-2、IL-4、IFN-γ和IL-10 mRNA。ST-SPM在48小时时细胞因子mRNA的重新表达增强，同时IL-2、IFN-γ、IL-4和IL-10蛋白的产生增加。这些结果表明，在初次免疫反应期间T细胞暴露于尼古丁而非ST会导致4天后细胞因子表达过度。此外，在初次免疫反应期间最初暴露于尼古丁或ST的记忆T细胞，在不存在尼古丁或ST的情况下再次刺激T细胞时会表现出过度的细胞因子表达。药理学。
