Journal: Molecular Therapy. Nucleic Acids

Article Title: mRNA-1273 is placenta-permeable and immunogenic in the fetus

doi: 10.1016/j.omtn.2025.102489

Figure Lengend Snippet: Anti-spike cellular immunity in dams and their pups after maternal mRNA-1273 vaccination during pregnancy (A and B) After maternal vaccination with 4 μg mRNA-1273 twice, the dams and pups were examined for spike-specific lymphocyte proliferation by the readout of incorporated tritium in vitro . Splenic lymphocytes of both dams ( p = 0.001) and pups ( p < 0.001) proliferated specifically in response to spike, as opposed to those with maternal saline injection. Besides, the dams ( p = 0.004) and pups ( p < 0.001) with maternal mRNA-1273 vaccination were superior in spike-specific lymphocyte proliferation to their respective saline controls. Bovine serum albumin (BSA) was the third-party stimulators, and Con-A was a mitogen to non-specifically stimulate T cells. (C and D) IFN- γ - and IL-2 ELISpot images in triplicate shown were from a representative dam and pup with maternal mRNA-1273 (4 μg) or saline vaccination during pregnancy. Both groups exhibited heightened frequencies of IFN- γ - and IL-2-secreting T cells, as compared with their respective control counterparts. Error bar charts display the boxed areas of 95% confidence intervals for the means as box-crossing horizontal lines.

Article Snippet: Murine IFN-γ/IL-2-secreting T cells were quantified by mouse IFN-γ and IL-2 ELISpot Kits according to the manufacturer’s instructions (R&D Systems).

Techniques: In Vitro, Saline, Injection, Enzyme-linked Immunospot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: mRNA-1273 is placenta-permeable and immunogenic in the fetus

doi: 10.1016/j.omtn.2025.102489

Figure Lengend Snippet: Immunological consequences of in utero mRNA-1273 injection GD14 FVB/N fetuses were subjected to intraperitoneal injection of mRNA-1273 (IU mRNA-1273, n = 19). (A and B) Postnatally, serum anti-spike IgG 1 /IgG 2a was examined at the age of 1 month. IU mRNA-1273 led to significantly higher titers of anti-spike IgG 1 /IgG 2a , as compared with in utero saline injection (IU saline, n = 9). Serum anti-spike IgG 1 /IgG 2a gradually decreased within postnatal 3 months. Circles interconnected by a line represent IgG 1 /IgG 2a levels measured at 1 (M1), 2 (M2), and 3 (M3) months old from an individual mouse ( n = 11). (C) Lymphocyte proliferation in response to spike protein was measured by the readout of incorporated tritium ( n = 4) as counts per minute (cpm). Medium only was used as background controls, BSA as third-party stimulators, and Con-A as a mitogen to stimulate the T cell population. IU mRNA-1273 significantly proliferated specifically in response to spike protein ( p < 0.027), whereas IU saline ( n = 4) failed to show lymphocyte proliferation under spike protein stimulation. There was a significant difference in lymphocyte proliferation under spike protein stimulation between IU mRNA-1273 and IU saline ( p < 0.006). Rectangles within a dataset represent 95% confidence intervals for the means, which are shown as transverse lines crossing the rectangles. (D) Spike-reactive IFN- γ - and IL-2-secreting cells of splenic lymphocytes were enumerated by ELISpot. Figures showed the spots with their counts from the representative mice of IU mRNA-1273 and IU saline. The frequency of spike-reactive IFN- γ - and IL-2-secreting T cells was calculated by the mean of ELISpot readouts (triplicates) divided by the CD3 + cell ratio of splenic lymphocytes in each individual mouse.

Article Snippet: Murine IFN-γ/IL-2-secreting T cells were quantified by mouse IFN-γ and IL-2 ELISpot Kits according to the manufacturer’s instructions (R&D Systems).

Techniques: In Utero, Injection, Saline, Enzyme-linked Immunospot

Journal: BMC Biotechnology

Article Title: Extracellular overproduction of E7 oncoprotein of Iranian human papillomavirus type 16 by genetically engineered Lactococcus lactis

doi: 10.1186/s12896-019-0499-5

Figure Lengend Snippet: Panel a : The levels of E7-specific IgG and SIgA antibodies in serum (left) and vaginal discharges (right), respectively, on day 41; the sera and vaginal discharge samples were tested by indirect ELISA using recombinant human papillomavirus type 16 protein E7 as the coating antigen. Goat-anti-mouse IgG-H&L (HRP) antibody and Goat Anti-Mouse IgA alpha chain (HRP) were used as a secondary antibody. The absorbance of each microwell was read on a spectrophotometer at 450 nm. Error bars represent the mean ± standard error value of each group ( P < 0.001). Panels b and c : Detection of E7-specific T-lymphocytes using an IL-2 (panel b ) and IFN-γ (panel c ) ELISPOT assay in the splenocytes (left) and intestinal mucosal lymphocytes (right); female C57BL/6 mice were immunized orally with 10 9 CFU of Lactococcus lactis containing rE7 on days 1, 2, 3, 14, 15, 16, 29, 30, and 31. In order to measure cytokine production from the cells, on day 41 (ten days after the last immunization), intestinal mucosal lymphocytes and splenocytes were collected and stimulated with MHC class I - restricted peptides and MHC class II - restricted peptides (HPV-16 E7 49–57 and HPV-16 E7 30–67 , respectively). Then, the level of E7-specific IFN-γ-producing CD8 + and CD4 + T cells and E7-specific IL-2-producing CD8 + and CD4 + T cells was calculated via mouse IFN-γ and IL-2 ELISPOT kit, respectively. The diagram demonstrates the number of spots per 10 4 splenocytes and intestinal mucosal lymphocytes after stimulation. The results are presented as mean ± SD (n 10). PBS (Control group); LLEV: Induced L. lactis harboring pNZ8123; LLE7BF: Induced L. lactis harboring pNZ8123-HPV16-E7 produced through batch fermentation; LLE7FBF: Induced L. lactis harboring pNZ8123-HPV16-E7 produced through fed-batch fermentation; LLOE7BF: Induced L. lactis harboring pNZ8123-HPV16-optiE7 produced through batch fermentation; and LLOE7FBF: Induced L. lactis harboring pNZ8123-HPV16-optiE7 produced through fed-batch fermentation

Article Snippet: Afterwards, the number of E7-specific IFN-γ-producing CD8 + and CD4 + T cells and E7-specific IL-2-producing CD8 + and CD4 + T cells were measured using mouse IFN-γ ELISPOT kit and mouse IL-2 mouse ELISPOT kit, respectively, according to the manufacturer’s instructions (R&D Systems, Inc., Minneapolis, MN).

Techniques: Indirect ELISA, Recombinant, Spectrophotometry, Enzyme-linked Immunospot, Control, Produced