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To assess both benefits and risks related to treatment interruption (TI) in children with chronic HIV-1 materno-fetal infection.A multicentre, retrospective analysis in five university hospital pediatric departments in France.Clinical events, plasma HIV-1 RNA, CD4 cell counts, CD4 percentages (CD4%) and genotypes were recorded in 24 patients before and during TI. Patients were classified as sparing regimen or virological failure groups according to the main reason for treatment interruption. Clinical events, immuno-virological evolution and genotype reversions were monitored.After a median of 40 weeks of TI, none of the patients presented with an AIDS-defining event. For the whole cohort, median viral load variation from baseline, measured during TI was +1.26 log10 copies/ml (range, −0.22, +4.3 log10) with large inter-individual variability, median absolute CD4 cell loss was 32.5% (range, −82, +17%). These variations were not different in the two patient groups. The mean number of mutations conferring resistance to nucleoside reverse transcriptase inhibitors, non-nucleoside reverse transcriptase inhibitors and protease inhibitors at baseline and last evaluation did not differ significantly. Few mutation reversions to wild type were noted in our cohort.Treatment interruption in children with chronic HIV-1 infection is associated with higher viral load increases than observed in adult patients. The CD4 cell loss is comparable. Although no clinical AIDS-defining event was noted close monitoring is required when TI is proposed to HIV-infected children. Very few reversion mutations were observed during treatment interruption.