Isolation of Mononuclear Cells from Whole Blood: Comparison of Two Media

Zito, Denise R.; Johnson, Kristen; Skipper, Mauver E.; Normansell, David E.

doi:10.1093/labmed/17.2.94

Cited by 1 publication

(2 citation statements)

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“…Details for the isolation of mononuclear cells and culture conditions for lymphocyte proliferation have been previously described. 19,27 The culture medium contained RPMI-1640, 10% heat-inactivated human AB serum, 5 × 10 4 units penicillin, 50 mg streptomycin, 1 mmol sodium pyruvate, 0.1 mmol nonessential amino acids, 2 mmol L-glutamine, and 50 μmol β-mercaptoethanol (per 1,000 mL). Lymphocyte proliferation (of triplicate cultures containing 2 × 10 5 viable cells/200 μL medium) was assessed by the rate of 3 H-thymidine incorporated into DNA.…”

Section: Lymphocyte Proliferationmentioning

confidence: 99%

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In Vitro Lymphocyte Functions in Undernourished Children With Sickle Cell Anemia

Kuvibidila

Gardner

Velez

et al. 2020

TOJ

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Background: Children with sickle cell disease (SCD) often suffer from growth deficits and impaired immunity. However, the association between mild to moderate malnutrition and in vitro lymphocyte function has not been well studied. The goal of this study was to investigate the effects of undernutrition on lymphocyte functions in children with SCD. Methods: Weight; height; plasma concentrations of albumin (Alb), prealbumin (PA), transferrin (Tf), retinol-binding protein (RBP), α 1-acid glycoprotein (AGP), C-reactive protein (CRP), and ceruloplasmin (Cp); and lymphocyte proliferation and interleukin (IL)-2 in phytohemagglutinin-treated blood lymphocytes were measured in 90 children with SCD (59 SS, 4 Sβ°, 27 SC hemoglobin genotypes). Results: The mean age of the children included in the analysis was 7.65 years. Four of the 90 children had weight and height below the fifth percentile. A higher percentage of children with HbSS/HbSβ°(61.4%) than of those with HbSC (44%) had ࣙ2 plasma protein concentrations below normal (Alb <35 g/L, PA <160 mg/L, Tf <2.0 g/L, and RBP ࣘ20 mg/L). Mean anthropometric measurements, hemoglobin, and hematocrit were lower in children with HbSS/HbSβ°than in those with HbSC (P<0.05). Lymphocyte proliferation was reduced by 20% to 27% in children with HbSS/HbSβ°with undernutrition plus inflammation (AGP >1 g/L, CRP >5 mg/L, Cp >600 mg/L) compared to children with neither. Regardless of inflammatory status, lymphocyte proliferation was reduced by 29% to 49% in children with HbSS/HbSβ°and undernutrition defined by PA or Alb plus RBP (P<0.05) compared to those with RBP within normal range. Neither undernutrition nor inflammation reduced lymphocyte proliferation in children with HbSC. Mean IL-2 activity was reduced by undernutrition, defined as PA <160 mg/L, in both groups. PA, RBP, and hemoglobin concentrations positively correlated with in vitro lymphocyte functions (P<0.05). Conclusion: Undernutrition altered in vitro lymphocyte function in children with the HbSS/HbSβ°genotypes. Dietary supplements may improve the altered functions in these children.

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Section: Lymphocyte Proliferationmentioning

confidence: 99%

“…The IL-2 activity in the supernatant was estimated by the growth rate of IL-2-dependent CTLL-2 cells assessed by 3 H-thymidine uptake. 27 In the absence of IL-2, CTLL-2 cells (which are derived from murine cytotoxic T cells) do not undergo cell multiplication. 28…”

Section: Interleukin-2 Studymentioning

confidence: 99%