Project description:Class-switching to IgG2a/c in mice is a hallmark response to intracellular pathogens. T cells can promote class-switching and the predominant pathway for induction of IgG2a/c antibody responses has been suggested to be via stimulation from Th1 cells. We previously formulated CAF®01 (cationic liposomes containing dimethyldioctadecylammonium bromide (DDA) and Trehalose-6,6-dibehenate (TDB)) with the lipidated TLR7/8 agonist 3M-052 (DDA/TDB/3M-052), which promoted robust Th1 immunity in newborn mice. When testing this adjuvant in adult mice using the recombinant Chlamydia trachomatis (C.t.) vaccine antigen CTH522, it similarly enhanced IgG2a/c responses compared to DDA/TDB, but surprisingly reduced the magnitude of the IFN-g+ Th1 response in a TLR7 agonist dose-dependent manner. Single cell RNA-sequencing revealed that DDA/TDB/3M-052 liposomes initiated early transcription of class-switch regulating genes directly in pre-germinal center B cells. Mixed bone marrow chimeras further demonstrated that this adjuvant did not require Th1 cells for IgG2a/c switching, but rather facilitated TLR7-dependent T-bet programming directly in B cells. This study underlines that adjuvant-directed IgG2a/c class-switching in vivo can occur in the absence of T cell help, via direct activation of TLR7 on B cells and positions DDA/TDB/3M-052 as a powerful adjuvant capable of eliciting type I-like immunity in B cells without strong induction of Th1 responses.
Project description:Untargeted-metabolomics LC-MS/MS analysis of commercial natural products pool, analyzed with different DDA settings with the objective to find the best one.
Project description:Untargeted-metabolomics LC-MS/MS analysis of commercial natural products pool, analyzed with different DDA settings with the objective to find the best one.
Project description:Results of an unbiased series of 2833 fresh products of conception samples (in conjunction with maternal blood samples for comparison) that were tested using a 300K Illumina SNP array. we report on the rate and type of whole aneuploidies, UPD, partial aneuploidies, copy number variants found in our series as well as the rate of maternal cell contamination vs. true fetal results. 2833 consecutive fresh products of conception samples and corresponding maternal and/or paternal blood samples were analyzed using a SNP array plus informatics algorithms for the purpose of detecting causal chromosome abnormalities. Hypothesized advantages of using SNP microarray with Parental SupportTM informatics over the traditional standard G-banded karyotyping include: direct testing without need for cell culture, faster turn-around time, low failure rate, the ability to detect or rule out maternal cell contamination (MCC), and the detection of small segmental changes, uniparental disomy (UPD) and parent of origin of any aneuploidies. Results showed a 22% MCC rate. In the 78% of samples with true fetal results, 60% had abnormalities with single aneuploidy being the most common. Separate evaluation of the samples to determine the resolution of the SNP array with informatics showed the ability to detect copy number variations (CNVs) as small as 1 MB or less, with 1.4% of samples revealing a clinically relevant microdeletion or duplication. 2833 fresh products of conception samples were analyze using no controls, no replicates, and no reference samples.