The Effect Of Glutamine On Some Immune Character Of Peripheral Blood Mononuclear (PBMCs) From Neonate

Objective To investigate the effect of glutamine on the proliferatiofi andactivation of peripheral blood mononuclear cells (PBMCs) from different neonates, and the differentiation of T lymphocyte subsets and the activity of NK cells from PMBCs ate also analyzed.Methods PBMCs from premature neonates, term neonates and infectiveneonates were cultured respectively with or without glutamine. 1. PBMCs proliferation assay: The proliferation of PBMCs stimulated by T-cell mitogen PHA for 48 hours was determined by means of the 3-(4-5-dimethylthiazol) 2,5-diphenyl tetrazolium bromide (MTT) assay. 2.The expression of the early activation marker of PBMCs: Applying reverse transcription-polymerase chain reaction (RT-PCR) technique to detect IL-2 mRNA expression of PBMCs stimulated by T-cell mitogen PHA for 48 hours .3.The expression of the late activation marker of PBMCs: Applying flow-cytometry (FCM) to analyze CD25 expression on PBMCs stimulated by T-cell mitogen ConA for 66-72 hours .4 The differentiation of T lymphocytes: Applying FCM to analyze CD4 and CD8 expression on PBMCs stimulated by T-cell mitogen ConA for 66-72 hours. 5. The activity of NK cells: Applying FCM to analyze CD3/CD16+CD56 expression on PBMCs stimulated by IL-2 and LPS for 96 hours.Results 1. PBMC proliferation assay: Glutamine increased the prolifeation ofT-cell mitogen- induced PBMCs from neonates. 2.IL-2 mRNA expression: IL-2 mRNA expression of PBMC from different neonates was not affected by glutamine.3. CD25 expression: Glutamine significantly increased Positive Expression Cell Rate (PECR) of CD25 on ConA- stimulated PBMCs from premature neonates and term neonates,AbstractMean Fluorescence Intensity (RMFI) of CD25 on ConA- stimulated PBMC of term neonates also increased by glutamine. But glutamine did not influence CD25 expression on ConA- stimulated PBMC of infective neonates and RMFI of CD25 on ConA- stimulated PBMC of premature neonates.4.The differentiation of T lymphocytes: Glutamine significantly increased the proportion and counts of CD4+ lymphocytes and improved the ratio of CD4:CD8 lymphocytes from neonates after ConA stimulation. 5. The activity of NK cells: Glutamine did not influence PECR of CD3-CD16+CD56+ on PBMCs simulated by IL-2 and LPS for 96 hours from premature neonates and term neonates, but glutamine significantly increased PECR of CD3-CD16+CD56+ on PBMCs of infective neonates.Conclusions 1 .Glutamine promotes the proliferation of PBMCs from differentneonates 2. Glutamine enhances the activation of neonatal PBMCs.3. Glutamint can increase the proportion and counts of CD4+ lymphocytes and improve the ratio of CD4+: CDS +lymphocytes from neonates. It suggests that glutamine could promote the maturation and differentiation of neonatal T lymphocytes. 4.Glutamine could recover the activity of NK cell of infective neonates. It suggests that glutamine could ameliorate immunosuppressive state of neonates when infected. So glutamine to some extent could promote the proliferation of neonatal PBMC and the maturation of neonatal immunity.