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In vitro IgE synthesis in neonates with different family history of atopy
S Miceli Sopo1, M A Pesaresi, G Minchilli
1Department of Pediatrics, Catholic University of Rome.
Insights
Neonatal blood mononuclear cells (NBMC) show limited in vitro synthesis of IgE, IL4, and IFN gamma, regardless of family history of atopy. These tests are not reliable markers for predicting atopy predisposition in newborns.
Area of Science:
- Immunology
- Neonatal Research
- Allergy and Atopy Studies
Background:
- Atopy, a genetic predisposition to allergic diseases, is often assessed through family history.
- Understanding early immune responses in neonates is crucial for predicting atopy development.
- In vitro immune cell function assays may offer insights into atopy risk.
Purpose of the Study:
- To investigate the in vitro synthesis of IgE, IL4, and IFN gamma by neonatal blood mononuclear cells (NBMC).
- To correlate NBMC cytokine production and IgE synthesis with family history of atopy (biparental, uniparental, or absent).
- To evaluate the utility of these in vitro tests as potential markers for atopy predisposition.
Main Methods:
- NBMC from neonates with varying family histories of atopy were stimulated with PHA and TPA.
- Lymphokine production (IL4, IFN gamma) was measured via ELISA.
- NBMC were pre-incubated with rhIL4, then stimulated under various conditions to assess IgE synthesis.
Main Results:
- Only one neonate showed detectable IL4 production; no significant IFN gamma was detected in any group.
- Spontaneous and rhIL4-stimulated IgE production by NBMC was minimal and not significantly increased by other stimuli.
- No significant differences in cytokine or IgE production were observed among the three groups based on family history.
Conclusions:
- NBMC demonstrate limited capacity for significant in vitro IgE, IL4, or IFN gamma production.
- Current in vitro assays measuring IgE, IL4, and IFN gamma synthesis are not effective markers for predicting atopy predisposition in neonates.
- Family history alone may not fully predict early immune responses related to atopy.
Abstract:
In this study we want to correlate family history of atopy and in vitro synthesis of IgE, IL4 and IFN gamma in 5 neonates with biparental (group A), 5 with uniparental (group B) and 5 with absent family history of atopy (group C). An aliquot of neonatal blood mononuclear cells (NBMC) was incubated in presence of PHA in combination with the phorbol ester acetate (TPA). The supernatants of cultures were harvested after 48-72 hours of incubation and stored at -20 degrees C until testing for lymphokine production by ELISA kits. Only one neonate of group B showed detectable in vitro synthesis of IL4 (45 pg/ml) after PHA + TPA stimulation. All the others failed to produce detectable levels either of IL4 or IFN gamma. Another aliquot of NBMC was cultured in the presence of saturating concentrations of rhIL4 for 48 h. After this pre-incubation step, the non-adherent cells were cultured in the presence of: 1) rhIL4; 2) rhIL4 + anti-IL4 antibody (Ab); 3) rhIL4 + anti-IFN gamma Ab; 4) rhIFN gamma, 5) rhIFN gamma + anti-IFN gamma Ab + rhIL4; 6) PWM + rhIL4; 7) unstimulated culture. The supernatants of these cultures were tested for their IgE content. In general, spontaneous IgE production by NBMC was very low, rhIL4 did not induce a significant increase of IgE synthesis. The other modalities of stimulations did not produce significative changes. We did not observe significative differences among the three groups of neonates. On the basis of our results, we conclude that NBMC aren't able to produce significative amounts of IgE in vitro either spontaneously or after IL4 stimulation. This test and the evaluation of IL4 and IFN gamma in vitro production aren't useful markers of atopy predisposition.