hCD8A

Fig.1 Detection of CD8A expression in thymus by RT-PCR. 

Wild type: only one band at 248 bp with primers F1/R1(mCd8a);

Homozygous: only one band at 124 bp with primers F2/R2(hCD8A).

Abbr. M, DNA marker; HO, homozygous; WT, wild type.

Fig.2 Detection of hCD8A expression in Blood and Spleen in hCD8A and WT mice.

Fig.3 Detection of immune cell subpopulations in the blood by FACS (n=3 in all groups). 

Abbr.  WT, wild type; HO, homozygous.

Fig.4 Detection of immune cell subpopulations in the spleen by FACS (n=3 in all groups). 

Abbr.  WT, wild type; HO, homozygous.

Fig.5 Body weight and body weight change among all groups. (n=3, 7 weeks-old, male). Mice treated with anti-hCD8 VHH-LNP exhibited no significant decreased absolute body weight.

Fig.6 In vivo bioluminescence imaging of hCD8A-targeted LNP in HO hCD8A mice. WT C57BL/6 and HO hCD8A mice (n=3, 7 weeks-old, male) were intravenously administered with either 10 µg or 30 µg of CD8a VHH‑conjugated LNP encapsulating luciferase‑encoding eGFP mRNA. (A)Whole body image from dorsal and ventral side were imaged 2-day post-injection; (B) IVIS signal was quantified, and the standard error of the mean±SEM was calculated. 

In HO hCD8A mice, the anti-hCD8 VHH-LNP mainly delivered mRNA to the liver (chest region).

Fig.7 In vivo bioluminescence imaging of organs in HO hCD8A mice treated with hCD8A-targeted LNP. WT C57BL/6 and HO hCD8A mice (n=3, 7 weeks-old, male)were intravenously administered with either 10 µg or 30 µg of anti-hCD8a VHH-conjugated LNP encapsulating luciferase mRNA. Organs including brain, heart, liver, spleen, lung, kidney, and lymph nodes (LN) were harvested and luciferase expression was assessed by IVIS. (A) Representative bioluminescence signals (in relative radiance, e.g., photons/sec/cm²/sr) are shown for each organ across different dose groups. (B) IVIS signal was quantified, and SEM was calculated. 

CD8a-tLNP showed dose-dependent targeting and superior spleen delivery in HO hCD8A mice compared to WT C57BL/6 mice.