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Effective boosting of both Gag-specific CD8 and CD4 T cell responses was observed at one week following the second immunization (1st booster immunization) in both groups of immunized macaques, but was significantly more pronounced (group median responses were approximately 2-fold higher) in those macaques that were immunized with the MVAΔudg-gag vector as compared to MVA-gag (Figure 9A, B, D).
ELISPOT analyses indicated that the average Gag-specific IFN-γ response elicited by the hLAMP/gag chimera was detectable after only two or three naked DNA immunizations in all five immunized macaques and reached an average of 1000 spot-forming cells (SFC /106 PBMCs.
The responses included highly significant IgG antibody titers after two DNA immunizations in four of the five immunized macaques and IgA responses in serum and nasal washes.
Similar findings have been obtained with the five immunized macaques, each of which showed strong humoral immune responses (Figure 8).
Among them, the attenuated strains of vaccinia virus MVA and NYVAC have been considered as promising HIV/AIDS vaccine candidates due to their strong safety record in preclinical and clinical trials, high stability, ability to induce potent and long-lasting immune responses to foreign antigens, and efficacy in immunized macaques after challenge with SIV [15].
More importantly, these Δ5G "immunized" macaques during the chronic phase manifested near-sterile immunity when challenged with the homologous wild-type SIVmac239, and the animals showed neither evolution of pathogenic revertants nor clinical disease manifestation during a 10 year follow up period.
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Analysis of memory B cells from the immunized macaque suggests that elicitation of broadly neutralizing antibodies (bNAbs) for ebolaviruses is possible but difficult, potentially due to the rarity of bNAb clones and their precursors.
Six and nine days respectively after the last boost, RNA was isolated using Tri Reagent (Molecular Research Center Inc ,Cincinnati, USA) from the bone marrow of the immunized macaque and transferred into cDNA by reverse transcription.
For each antigen, a responder Rhesus macaque was defined as one exhibiting a significant number IFN-γ SFC cells over the background value or value for non-immunized macaques, at any time point over the immunization course.
In contrast, among the four ID-immunized macaques, two presented multi-specific T-cell responses to as many as four pools of SIV-NEF and/or GAG peptides.
We find that immunization of rhesus macaques with the pentavalent vaccine results in protection of 55% of pentavalent-vaccine-immunized macaques from simian-human immunodeficiency virus (SHIV) challenge.
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