Understanding the mechanisms underlying autoantibody development will accelerate therapeutic target identification in autoimmune diseases such as Systemic Lupus Erythematosus (SLE) 1 . Follicular helper T cells (Tfh) have long been implicated in SLE pathogenesis. Yet, a fraction of SLE patients’ autoantibodies are unmutated, supporting that autoreactive B cells also differentiate outside germinal centers (GCs) 2 . Here, we describe a CXCR5 − CXCR3 + PD1 hi CD4 + T cell helper population distinct from Tfh and expanded in both SLE blood and the tubulointerstitial areas of patients with Proliferative Lupus Nephritis (PLN). These cells produce IL10 and accumulate mitochondrial ROS (mtROS) as the result of reverse electron transport (RET) fueled by succinate. Furthermore, they provide B cell help, independently of IL21, through IL10 and succinate. Similar cells are generated in vitro upon priming naïve CD4 + T cells with plasmacytoid DCs (pDCs) activated with Oxidized mitochondrial DNA (Ox mtDNA), a distinct class of interferogenic TLR9 ligand 3 . Targetting this pathway might blunt the initiation and/or perpetuation of extrafollicular humoral responses in SLE.

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Immunization with HIV AIDSVAX gp120 vaccines in the phase III VAX003 and VAX004 trials did not confer protection. To understand the shortcomings in antibody (Ab) responses induced by these vaccines, we evaluated the kinetics of Ab responses to the V1V2 and V3 regions of gp120 and the induction of Ab-mediated antiviral functions during the course of 7 vaccinations over a 30.5-month period. Plasma samples from VAX003 and VAX004 vaccinees and placebo recipients were measured for ELISA-binding Abs and for virus neutralization, Ab-dependent cellular phagocytosis (ADCP), and Ab-dependent cellular cytotoxicity (ADCC). Ab responses to V1V2 and V3 peaked after 3 to 4 immunizations and declined after 5 to 7 immunizations. The deteriorating responses were most evident against epitopes in the underside of the V1V2 β-barrel and in the V3 crown. Correspondingly, vaccinees demonstrated higher neutralization against SF162 pseudovirus sensitive to anti-V1V2 and anti-V3 Abs after 3 or 4 immunizations than after 7 immunizations. Higher levels of ADCP and ADCC were also observed at early or mid-time points as compared with the final time point. Hence, VAX003 and VAX004 vaccinees generated V1V2- and V3-binding Abs and functional Abs after 3 to 4 immunizations, but subsequent boosts did not maintain these responses.

The V3 loop in the HIV envelope gp120 is one of the immunogenic sites targeted by Abs. The V3 crown in particular has conserved structural elements recognized by cross-reactive neutralizing Abs, indicating its potential contribution in protection against HIV. Crystallographic analyses of anti-V3 crown mAbs in complex with the V3 peptides have revealed that these mAbs recognize the conserved sites on the V3 crown via two distinct strategies: a cradle-binding mode (V3C) and a ladle-binding (V3L) mode. However, almost all of the anti-V3 crown mAbs studied in the past were isolated from chronically HIV-infected individuals. The extents to which the two types of anti-V3 crown Abs are generated by vaccination are unknown. This study analyzed the prevalence of V3C-type and V3L-type Ab responses in HIV-infected individuals and in HIV envelope-immunized humans and animals using peptide mimotopes that distinguish the two Ab types. The results show that both V3L-type and V3C-type Abs were generated by the vast majority of chronically HIV-infected humans, although the V3L-type were more prevalent. In contrast, only one of the two V3 Ab types was elicited in vaccinated humans or animal models, irrespective of HIV-1 envelope clades, envelope constructs (oligomeric or monomeric), and protocols (DNA plus protein or protein alone) used for vaccinations. The V3C-type Abs were produced by vaccinated humans, macaques, and rabbits, whereas the V3L-type Abs were made by mice. The V3C-type and V3L-type Abs generated by the vaccinations were able to mediate virus neutralization. These data indicate the restricted repertoires and the species-specific differences in the functional V3-specific Ab responses induced by the HIV envelope vaccines. The study implies the need for improving immunogen designs and vaccination strategies to broaden the diversity of Abs in order to target the different conserved epitopes in the V3 loop and, by extension, in the entire HIV envelope.

The V3 region of HIV-1 gp120 is important for virus-coreceptor interaction and highly immunogenic. Although most anti-V3 antibodies neutralize only the sensitive tier 1 viruses, anti-V3 antibodies effective against the more resistant viruses exist, and a better understanding of these antibodies and their epitopes would be beneficial for the development of novel vaccine immunogens against HIV. The HIV-1 isolate JRFL with its cryptic V3 is resistant to most V3-specific monoclonal antibodies (MAbs). However, the V3 MAb 2424 achieves 100% neutralization against JRFL. 2424 is encoded by IGHV3-53 and IGLV2-28 genes, a pairing rarely used by the other V3 MAbs. 2424 also has distinct binding and neutralization profiles. Studies of 2424-mediated neutralization of JRFL produced with a mannosidase inhibitor further revealed that its neutralizing activity is unaffected by the glycan composition of the virus envelope. To understand the distinct activity of 2424, we determined the crystal structure of 2424 Fab in complex with a JRFL V3 peptide and showed that the 2424 epitope is located at the tip of the V3 crown ( IMPORTANCEHIV/AIDS vaccines are crucial for controlling the HIV epidemics that continue to afflict millions of people worldwide. However, HIV vaccine development has been hampered by significant scientific challenges, one of which is the inability of HIV vaccine candidates evaluated thus far to elicit production of potent and broadly neutralizing antibodies. The V3 loop is one of the few immunogenic targets on the virus envelope glycoprotein that can induce neutralizing antibodies, but in many viruses, parts of V3 are inaccessible for antibody recognition. This study examined a V3-specific monoclonal antibody that can completely neutralize HIV-1 JRFL, a virus isolate resistant to most V3 antibodies. Our data reveal that this antibody recognizes the most distal tip of V3, which is not as occluded as other parts of V3. Hence, the epitope of 2424 is in one of the vulnerable sites on the virus that may be exploited in designing HIV vaccine immunogens. The HIV-1 envelope glycoprotein (Env) is the only virus-encoded protein expressed on the surface of the virus and is the sole target for virus-neutralizing antibodies (Abs). On the virion surface, the HIV Env spike is a compact heterodimeric trimer made up of gp120 and gp41 subunits (1-3). The surface gp120 subunit is responsible for interacting with the host cell through binding to CD4 and the coreceptor, the chemokine receptor CCR5 or CXCR4 (4-7). On the basis of primary amino acid sequences, gp120 is divided into five conserved regions (C1 to C5), which are interspersed with five variable regions (V1 to V5) (8). The CD4-binding site and the chemokine receptor-binding site are both highly conformational and discontinuous. The chemokine receptor binding site in particular is composed of the invariant ␤2 and ␤3 strands of the V1V2 stem region, ␤20 and ␤21 strands in the conserved C4 region, and the third variable (V3) region of gp120 (3, 9). Vulnerable sites on t...