HIV-specific humoral immune responses by CRISPR/Cas9-edited B cells

Hartweger, Harald; McGuire, Andrew T.; Horning, Marcel; Jj, Taylor; Dosenovic, Pia; Yost, Daniel; Gazumyan, Anna; Seaman, Michael S.; Stamatatos, Leonidas; Janković, Mila; Nussenzweig, Michel C.

doi:10.1084/jem.20190287

Cited by 93 publications

(90 citation statements)

References 48 publications

(84 reference statements)

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“…As such, engineered BCRs can undergo class switching for eventual secretion as protective antibodies from plasma cells. B cells engineered in this way have been shown to confer protective levels of pathogen-specific antibody in vivo for several weeks following adoptive transfer into immunocompromised hosts 5 , or for several days following transfer into immunocompetent hosts 4,5 .…”

Section: Figurementioning

confidence: 99%

Vaccine Elicitation of HIV Broadly Neutralizing Antibodies from Engineered B cells

Huang

Tran

Olson

et al. 2020

Preprint

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24HIV broadly neutralizing antibodies (bnAbs) can suppress viremia and protect against 25 chronically infected patients shows that they generally derive from B cell precursors with uncommon 42 antigen receptor features such as long third heavy chain complementarity determining regions 43 (CDRH3s) which then require extensive somatic hypermutation 8,9 . 44 45 The gene sequences of HIV bnAbs and other desirable antibodies can however be engineered into 46 the genomes of ex vivo activated primary B cells, such that they are expressed as functional B cell 47 antigen receptors (BCRs) using endogenous heavy chain (HC) constant genes 3-5 . As such, 48 engineered BCRs can undergo class switching for eventual secretion as protective antibodies from 49 plasma cells. B cells engineered in this way have been shown to confer protective levels of pathogen-50 specific antibody in vivo for several weeks following adoptive transfer into immunocompromised 51 hosts 5 , or for several days following transfer into immunocompetent hosts 4,5 . 52 53Long-term expression of HIV bnAbs generated from engineered B cells, which could be boosted 54 through vaccination and which could mature in affinity against relevant viral sequences in 55 immunocompetent hosts, has potential as an attractive functional HIV cure strategy, given that 56 bnAbs administered passively in the context of infection have been shown to suppress viremia, kill 57 infected cells, and enhance host immunity 1,10,11 . Wild-type (WT) black 6 (C57BL/6J) mice represent 58 a useful model in which such an engineered B cell vaccine could be developed because of 59 similarities between mouse and human humoral immune systems, and because the mouse antibody 60 repertoire is also strongly genetically restricted in its ability to elicit HIV bnAbs [12][13][14] . In activated 61 primary B cells, we used CRISPR-Cas9 to insert the VRC01 HIV bnAb 15,16 light chain (LC) and HC 62 variable region (VDJ) into the mouse HC locus at J4 using a homology directed repair (HDR) genome 63 editing strategy. The inserted VRC01 gene is expressed under the control of a HC V-gene promoter 64 as a single mRNA, which is post-transcriptionally spliced to endogenous HC constant genes. A P2A 65 self-cleaving peptide sequence downstream of the mouse kappa (k) constant gene separates the 66 VRC01 light and heavy chains, allowing them to pair and form a functional cell surface expressed 67 BCR (H-targeting) (Fig. 1a). We chose to use the VRC01 genes because this prototype CD4 binding-68 site bnAb, which blocks entry of the virus into target CD4 + T cells, has been extensively tested in the 69 clinic for its ability to suppress viremia in patients and prevent infection after administration as a 70 recombinant monoclonal antibody 17-19 (clinicaltrials.gov NCT02568215, NCT02716675). 71 72A second strategy was also employed, in which the VRC01 heavy and kappa variable genes were 73 targeted separately to their endogenous loci for expression from V-gene promoter-controlled 74 transcripts spliced to cell-native ...

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Section: Figurementioning

confidence: 99%

Vaccine Elicitation of HIV Broadly Neutralizing Antibodies from Engineered B cells

Huang

Tran

Olson

et al. 2020

Preprint

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“…The avidity effect of nanoparticle-presentation provided an even stronger advantage to the B cell lineages that targeted crossneutralizing epitopes on the co-displayed RBDs, which led to broader NAb responses. Other recently developed or future technologies that we have not discussed, such as CRISPR/Cas9 genetic editing to produce bNAb-producing B cells for transplantation [274][275][276][277] or the use of osmotic pumps for the slow delivery of Env immunogens [278], might also be key for the development of the long-sought HIV-1 vaccine.…”

Section: Expert Opinionmentioning

confidence: 99%

Strategies for inducing effective neutralizing antibody responses against HIV-1

Moral-Sánchez

Sliepen

2019

Expert Review of Vaccines

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Introduction: Despite intensive research efforts, there is still no effective prophylactic vaccine available against HIV-1. Currently, substantial efforts are devoted to the development of vaccines aimed at inducing broadly neutralizing antibodies (bNAbs), which are capable of neutralizing most HIV-1 strains. All bNAbs target the HIV-1 envelope glycoprotein (Env), but Env immunizations usually only induce neutralizing antibodies (NAbs) against the sequence-matched virus and not against other strains. Areas covered: We describe the different strategies that have been explored to improve the breadth and potency of anti-HIV-1 NAb responses. The discussed strategies include the application of engineered Env immunogens, optimization of (bNAb) epitopes, different cocktail and sequential vaccination strategies, nanoparticles and nucleic acid-based vaccines. Expert opinion: A combination of the strategies described in this review and future approaches are probably needed to develop an effective HIV-1 vaccine that can induce broad, potent and long-lasting NAb responses.

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“…Like disease modelling, bloodderived iPSCs could present a convenient cell source for the production of various cell types of therapeutic values, such as endothelial cells, T cells and B cells. Since genetically modified B cells have been developed to generate anti-HIV-1 neutralizing antibodies (bNAbs) [48], clinically potent monoclonal B cells derived from iPSCs are considered to be a viable source of such antibodies. Antigenspecific T cells have excellent potential to alleviate the condition when injected to patients with cancer or virus infection.…”

Section: Therapeutic Applications Of Blood Reprogrammingmentioning

confidence: 99%

“…Apart from T cells, B-cell derivation could also be beneficial. Since genetically modified B cells have been developed to generate anti-HIV-1 neutralizing antibodies (bNAbs) [48], clinically potent monoclonal B cells derived from iPSCs are considered to be a viable source of such antibodies. More recently, iPSCs with enhanced immune-evading capacity have been established [49,50].…”

Section: Therapeutic Applications Of Blood Reprogrammingmentioning

confidence: 99%

Re‐entering the pluripotent state from blood lineage: promises and pitfalls of blood reprogramming

Chen

Yao

et al. 2019

FEBS Letters

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Edited by Catherine RobinBlood reprogramming, in which induced pluripotent stem cells (iPSCs) are derived from haematopoietic lineages, has rapidly advanced over the past decade. Since the first report using human blood, haematopoietic cell types from various sources, such as the peripheral bone marrow and cord blood, have been successfully reprogrammed. The volume of blood required has also decreased, from around tens of millilitres to a single finger-prick drop. Besides, while early studies were limited to reprogramming methods relying on viral integration, nonintegrating reprogramming systems for blood lineages have been subsequently established. Together, these improvements have made feasible the future clinical applications of blood-derived iPSCs. Here, we review the progress in blood reprogramming from various perspectives, including the starting materials and subsequent reprogramming strategies. We also discuss the downstream applications of blood-derived iPSCs, highlighting their clinical value in terms of disease modelling and therapeutic development.

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HIV-specific humoral immune responses by CRISPR/Cas9-edited B cells

Abstract: B cells edited by CRISPR/Cas9 to express anti–HIV-1 antibodies participate in humoral immune reactions after immunization and secrete neutralizing serum titers of anti-HIV antibodies in vivo.

Cited by 93 publications

References 48 publications

Vaccine Elicitation of HIV Broadly Neutralizing Antibodies from Engineered B cells

Vaccine Elicitation of HIV Broadly Neutralizing Antibodies from Engineered B cells

Strategies for inducing effective neutralizing antibody responses against HIV-1

Re‐entering the pluripotent state from blood lineage: promises and pitfalls of blood reprogramming

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