Language:

Identification of preexisting adaptive immunity to Cas9 proteins in humans

Charlesworth, Carsten T ; Deshpande, Priyanka S ; Dever, Daniel P ; Camarena, Joab ; Lemgart, Viktor T ; Cromer, M Kyle ; Vakulskas, Christopher A ; Collingwood, Michael A ; Zhang, Liyang ; Bode, Nicole M ; Behlke, Mark A ; Dejene, Beruh ; Cieniewicz, Brandon ; Romano, Rosa ; Lesch, Benjamin J ; Gomez-Ospina, Natalia ; Mantri, Sruthi ; Pavel-Dinu, Mara ; Weinberg, Kenneth I ; Porteus, Matthew H

Nature medicine, 2019-02, Vol.25 (2), p.249-254 [Peer Reviewed Journal]

United States: Nature Publishing Group

Full text available

Citations Cited by

Actions
1. Add to e-Shelf
2. Remove from e-Shelf
3. E-mail
4. Print
5. Permalink
6. Citation
7. EasyBib
8. EndNote
9. RefWorks
10. Delicious
11. Export RIS
12. Export BibTeX

Title:
Identification of preexisting adaptive immunity to Cas9 proteins in humans
Author: Charlesworth, Carsten T ; Deshpande, Priyanka S ; Dever, Daniel P ; Camarena, Joab ; Lemgart, Viktor T ; Cromer, M Kyle ; Vakulskas, Christopher A ; Collingwood, Michael A ; Zhang, Liyang ; Bode, Nicole M ; Behlke, Mark A ; Dejene, Beruh ; Cieniewicz, Brandon ; Romano, Rosa ; Lesch, Benjamin J ; Gomez-Ospina, Natalia ; Mantri, Sruthi ; Pavel-Dinu, Mara ; Weinberg, Kenneth I ; Porteus, Matthew H
Subjects: Adaptive Immunity ; Adaptive systems ; Adult ; Antibodies ; Antigens ; Care and treatment ; Cell Separation ; Clinical trials ; CRISPR ; CRISPR-Associated Protein 9 - metabolism ; Deoxyribonucleic acid ; DNA ; DNA sequencing ; Enzyme-linked immunosorbent assay ; Enzymes ; Female ; Gene editing ; Gene sequencing ; Genetic aspects ; Genomes ; Genomics ; Human populations ; Humans ; Immune response ; Immune response (cell-mediated) ; Immune response (humoral) ; Immunity ; Immunity, Humoral ; Inflammation ; Lymphocytes ; Lymphocytes T ; Male ; Medical research ; Nucleotide sequence ; Nucleotide sequencing ; Proteins ; Risk factors ; Staphylococcal infections ; Staphylococcus aureus ; Streptococcal infections ; T cells ; T-Lymphocytes - immunology
Is Part Of: Nature medicine, 2019-02, Vol.25 (2), p.249-254
Notes: ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
N.G.-O. and M.P.-D. aided in the design of immunoblot assays to detect Cas9. C.T.C. performed all immunoblots. J.C., V.T.L., and B.J.L. designed and performed an ELISA assay to detect antibodies against Cas9 in human serum. C.A.V., M.A.C., N.M.B., L. Z., and M.A.B. cloned, purified, and tested for purity both homologs of Cas9 used throughout the study. P.S.D., B.D., and M.K.C. designed and implemented the T cell assays and B.D. designed the flow cytometry panels. R.R., C.T.C., and B.C. designed and implemented the T cell expansion assays and restimulation assays. S.M. provided the cord blood serum. M.K.C., J.C., and C.T.C. performed the Ficoll-Paque density gradient to purify human PBMC. M.P-D. and P.S.D designed the figures. M.K.C., N.G.-O., C.T.C, M. H.P., and P.S.D. wrote the manuscript. K.I.W., D.P.D., and M.H.P. directed the research and participated in the design and interpretation of the experiments.
Author contributions
Description: The CRISPR-Cas9 system is a powerful tool for genome editing, which allows the precise modification of specific DNA sequences. Many efforts are underway to use the CRISPR-Cas9 system to therapeutically correct human genetic diseases . The most widely used orthologs of Cas9 are derived from Staphylococcus aureus and Streptococcus pyogenes . Given that these two bacterial species infect the human population at high frequencies , we hypothesized that humans may harbor preexisting adaptive immune responses to the Cas9 orthologs derived from these bacterial species, SaCas9 (S. aureus) and SpCas9 (S. pyogenes). By probing human serum for the presence of anti-Cas9 antibodies using an enzyme-linked immunosorbent assay, we detected antibodies against both SaCas9 and SpCas9 in 78% and 58% of donors, respectively. We also found anti-SaCas9 T cells in 78% and anti-SpCas9 T cells in 67% of donors, which demonstrates a high prevalence of antigen-specific T cells against both orthologs. We confirmed that these T cells were Cas9-specific by demonstrating a Cas9-specific cytokine response following isolation, expansion, and antigen restimulation. Together, these data demonstrate that there are preexisting humoral and cell-mediated adaptive immune responses to Cas9 in humans, a finding that should be taken into account as the CRISPR-Cas9 system moves toward clinical trials.
Publisher: United States: Nature Publishing Group
Language: English

Links

View this record in MEDLINE/PubMed

Back to results list

Previous Result 3 Next Go to Next page

Implemented by: Social Network Logos:

Identification of preexisting adaptive immunity to Cas9 proteins in humans

United States: Nature Publishing Group

Searching Remote Databases, Please Wait