Biomechanical Assessment of a Novel Sharp-Tipped Screw for 1-Step Minimally Invasive Pedicle Screw Placement Under Navigation.

BACKGROUND: The objective of this study was to assess the pullout force of a novel sharp-tipped screw developed for single-step, minimally invasive pedicle screw placement guided by neuronavigation compared with the pullout force for traditional screws. METHODS: A total of 60 human cadaveric lumbar pedicles were studied. Three different screw insertion techniques were compared: (A) Jamshidi needle and Kirschner wire without tapping; (B) Jamshidi needle and Kirschner wire with tapping; and (C) sharp-tipped screw insertion. Pullout tests were performed at a displacement rate of 10 mm/min recorded at 20 Hz. Mean values of these parameters were compared using paired t tests (left vs right in the same specimen): A vs B, A vs C, and B vs C. Additionally, 3 L1-L5 spine models were used for timing each screw insertion technique for a total of 10 screw insertions for each technique. Insertion times were compared using 1-way analysis of variance. RESULTS: The mean pullout force for insertion technique A was 1462.3 (597.5) N; for technique B, it was 1693.5 (805.0) N; and for technique C, it was 1319.0 (735.7) N. There was no statistically significant difference in pullout force between techniques (P > 0.08). The average insertion time for condition C was significantly less than that for conditions A and B (P < 0.001). CONCLUSIONS: The pullout force of the novel sharp-tipped screw placement technique is equivalent to that of traditional techniques. The sharp-tipped screw placement technique appears biomechanically viable and has the advantage of saving time during insertion. CLINICAL RELEVANCE: Single-step screw placement using high resolution 3-dimensional navigation has the potential to streamline workflow and reduce operative time.

Rod Attachment Induces Significant Strain in Lumbosacral Fixation.

STUDY DESIGN: This was a laboratory investigation. OBJECTIVE: Rod attachment can induce significant pedicle screw-and-rod pre- strain that may predispose the instrumentation to failure. This study investigated how in vitro L5-S1 rod strain and S1 screw strain during rod-screw attachment (pre-strain) compared with strains recorded during pure-moment bending ( test- strain). SUMMARY OF BACKGROUND DATA: The lumbosacral junction is highly vulnerable to construct failure due to rod fatigue fracture, sacral screw pull-out, and screw fatigue fracture. MATERIALS AND METHODS: Twelve cadaveric specimens were instrumented with L2-ilium pedicle screws and rod. Strain gauges on contoured rods and sacral screws recorded strains during sequential rod-to-screw tightening (pre-strains). The same instrumented constructs were immediately tested in a 6-degree-of-freedom apparatus under continuous loading to 7.5 Nm in multidirectional bending while recording instrumentation test-strains. Rod and screw pre-strains and test-strains were compared using 1-way repeated-measures analysis of variance followed by Holm-Sidák paired analysis (significant at P <0.05). RESULTS: The mean first (171±192 µE) and second (322±269 µE) rod attachment pre-strains were comparable to mean test-strains during flexion (265±109 µE) and extension (315±125 µE, P ≥0.13). The mean rod attachment pre-strain was significantly greater than mean test-strains during bidirectional lateral bending (40±32 µE ipsilateral and 39±32 µE contralateral, P <0.001) and axial rotation (72±60 µE ipsilateral and 60±57 µE contralateral, P <0.001). The mean first and second sacral screw pre-strains during rod attachment (1.03±0.66 and 1.39±1.00 Nm, respectively) did not differ significantly ( P =0.41); however, the mean sacral screw pre-strain during final (second) rod attachment was significantly greater than screw test-strains during all directions of movement (≤0.81 Nm, P ≤0.03). CONCLUSIONS: Instrumentation pre-strains imposed during in vitro rod-screw attachment of seemingly well-contoured rods in L2-ilium fixation are comparable to, and at times greater than, strains experienced during in vitro bending. Spine surgeons should be aware of the biomechanical consequences of rod contouring and attachment on construct vulnerability.

Influence of Spinal Deformity Construct Design on Adjacent-Segment Biomechanics.

BACKGROUND: Adjacent level degeneration is a precursor to construct failure in adult spinal deformity surgery, but whether construct design affects adjacent level degeneration risk remains unclear. Here we present a biomechanical profile of common deformity correction constructs and assess adjacent level biomechanics. METHODS: Standard nondestructive flexibility tests (7.5 Nm) were performed on 21 cadaveric specimens: 14 pedicle subtraction osteotomies (PSOs) and 7 anterior column realignment (ACR) constructs. The ranges of motion (ROM) at the adjacent free level in flexion, extension, axial rotation, and lateral bending were measured and analyzed. RESULTS: ACR constructs had a lower ROM change on flexion at the proximal adjacent free level than constructs with PSO (1.02 vs. 1.32, normalized to the intact specimen, P < 0.01). Lateral lumbar interbody fusion adjacent to PSO and 4 rods limits ROM at the free level more effectively than transforaminal interbody fusion and 2 rods in correction constructs with PSO. Use of 2 screws to anchor the ACR interbody further decreased ROM at the proximal adjacent free level on flexion, but adding 4 rods in this setting added no further limitation to adjacent segment motion. CONCLUSIONS: ACR constructs have less ROM change at the adjacent level compared to PSO constructs. Among constructs with ACR, anchoring the ACR interbody with 2 screws reduces motion at the proximal adjacent free level. When PSOs are used, lateral lumbar interbody fusion adjacent to the PSO level has a greater reduction in adjacent-segment motion than transforaminal interbody fusion, suggesting that deformity construct configuration influences proximal adjacent-segment biomechanics.

Biomechanics of a laterally placed sacroiliac joint fusion device supplemental to S2 alar-iliac fixation in a long-segment adult spinal deformity construct: a cadaveric study of stability and strain distribution.

OBJECTIVE: S2 alar-iliac (S2AI) screw fixation effectively enhances stability in long-segment constructs. Although S2AI fixation provides a single transarticular sacroiliac joint fixation (SIJF) point, additional fixation points may provide greater stability and attenuate screw and rod strain. The objectives of this study were to evaluate changes in stability and pedicle screw and rod strain with extended distal S2AI fixation and with supplemental bilateral integration of two sacroiliac joint fusion devices implanted using a traditional minimally invasive surgical approach. METHODS: Eight L1-pelvis human cadaveric specimens underwent pure moment (7.5 Nm) and compression (400 N) tests under 4 conditions: 1) intact (pure moment loading only); 2) L2-S1 pedicle screw and rod with L5-S1 interbody fusion; 3) added S2AI screws; and 4) added bilateral laterally placed SIJF. Range of motion (ROM), rod strain, and screw-bending moment (S1 and S2AI) were analyzed. RESULTS: Compared with S1 fixation, S2AI fixation significantly reduced L5-S1 ROM in right lateral bending by 50% (0.11°, p = 0.049) and in compression by 39% (0.22°, p = 0.003). Compared with fixation ending at S1, extending fixation with S2AI significantly decreased sacroiliac joint ROM by 52% (0.28°, p = 0.02) in flexion, by 65% (0.48°, p = 0.04) in extension, by 59% (0.76°, p = 0.02) in combined flexion-extension, and by 36% (0.09°, p = 0.02) in left axial rotation. The addition of S2AI screws reduced S1 screw-bending moment during flexion (0.106 Nm [43%], p = 0.046). With S2AI fixation, posterior L5-S1 primary rod strain increased by 124% (159 µE, p = 0.002) in flexion, by 149% (285 µE, p = 0.02) in left axial rotation, and by 99% (254 µE, p = 0.04) in right axial rotation. Compared with S2AI fixation, the addition of SIJF reduced L5-S1 strain during right axial rotation by 6% (28 µE, p = 0.04) and increased L5-S1 strain in extension by 6% (28 µE, p = 0.02). CONCLUSIONS: Long-segment constructs ending with S2AI screws created a more stable construct than those ending with S1 screws, reducing lumbosacral and sacroiliac joint motion and S1 screw-bending moment in flexion. These benefits, however, were paired with increased rod strain at the lumbosacral junction. The addition of SIJF to constructs ending at S2AI did not significantly change SI joint ROM or S1 screw bending and reduced S2AI screw bending in compression. SIJF further decreased L5-S1 rod strain in axial rotation and increased it in extension.

Biomechanics of open versus minimally invasive deformity correction:​ comparison of stability and rod strain between pedicle subtraction osteotomy and anterior column realignment.

OBJECTIVE: Anterior column realignment (ACR) is a new minimally invasive approach for deformity correction that achieves a degree of lordosis similar to that obtained with pedicle subtraction osteotomy (PSO). This study compared the biomechanical profiles of ACR with PSO using range of motion (ROM) and posterior rod strain (RS) to gain insight into the ACR technique and the necessary surgical strategies to optimize longevity and stability. METHODS: An in vitro biomechanical study using standard flexibility testing (7.5 Nm) was performed on 14 human cadaveric specimens, separated into 2 groups similar in age, sex, bone mineral density, and intact ROM. For group 1 (n = 7, instrumented L1-S1), a 30° ACR was performed at L3-4. For group 2 (n = 7, instrumented T12-S1), a 30° L3 PSO was performed. Specimens were subjected to nondestructive loads in flexion, extension, axial rotation, lateral bending, and compression. Conditions tested were 1) intact, 2) pedicle screw with 2 rods (PSR), 3) ACR or PSO with 2 rods (+2R), and 4) ACR or PSO with 4 rods (+4R). Primary outcome measures of interest were ROM stability and posterior RS at L3-4. RESULTS: No difference was observed between groups in lumbar lordosis (p = 0.83) or focal angular lordosis at L3-4 (p = 0.75). No differences in stability were observed between ACR+2R and PSO+2R (p ≥ 0.06);​ however, ACR+2R was significantly less stable than PSR in flexion and extension (p ≤ 0.02), whereas PSO+2R was less stable than PSR only in extension (p = 0.04). ACR+4R was more stable than ACR+2R in flexion, extension, left axial rotation, and compression (p ≤ 0.02). PSO+4R was more stable than PSO+2R only in extension (p = 0.04). Both ACR+2R and PSO+2R resulted in significant increases in RS in flexion and extension compared with PSR (p ≤ 0.032). RS in flexion and extension decreased significantly for ACR+4R versus ACR+2R and for PSO+4R versus PSO+2R (p ≤ 0.047). PSO+2R yielded lower RS than ACR+2R in compression (p = 0.03). No differences existed in RS between ACR+4R and PSO+4R (p ≥ 0.05). CONCLUSIONS: Although ACR appeared to be slightly more destabilizing than PSO using traditional 2R fixation, both techniques resulted in significant increases in posterior RS. The 4R technique increased stability in ACR and decreased RS in both ACR and PSO but may be more beneficial in ACR. Longer-term clinical studies are needed to appropriately identify the durability of the ACR technique in deformity correction.

Biomechanics of Circumferential Cervical Fixation Using Posterior Facet Cages: A Cadaveric Study.

OBJECTIVE: Anterior cervical discectomy and fusion (ACDF) is a common procedure for the treatment of cervical disease. Circumferential procedures are options for multilevel pathology. Potential complications of multilevel anterior procedures are dysphagia and pseudarthrosis, whereas potential complications of posterior surgery include development of cervical kyphosis and postoperative chronic neck pain. The addition of posterior cervical cages (PCCs) to multilevel ACDF is a minimally invasive option to perform circumferential fusion. This study evaluated the biomechanical performance of 3-level circumferential fusion with PCCs as supplemental fixation to anteriorly placed allografts, with and without anterior plate fixation. METHODS: Nondestructive flexibility tests (1.5 Nm) performed on 6 cervical C2-7 cadaveric specimens intact and after discectomy (C3-6) in 3 instrumented conditions: allograft with anterior plate (G+P), PCC with allograft and plate (PCC+G+P), and PCC with allograft alone (PCC+G). Range of motion (ROM) data were analyzed using 1-way repeated-measures analysis of variance. RESULTS: All instrumented conditions resulted in significantly reduced ROM at the 3 instrumented levels (C3-6) compared to intact spinal segments in flexion, extension, lateral bending, and axial rotation (p < 0.001). No significant difference in ROM was found between G+P and PCC+G+P conditions or between G+P and PCC+G conditions, indicating similar stability between these conditions in all directions of motion. CONCLUSION: All instrumented conditions resulted in considerable reduction in ROM. The added reduction in ROM through the addition of PCCs did not reach statistical significance. Circumferential fusion with anterior allograft, without plate and with PCCs, has comparable stability to ACDF with allograft and plate.

Variations Among Human Lumbar Spine Segments and Their Relationships to In Vitro Biomechanics: A Retrospective Analysis of 281 Motion Segments From 85 Cadaveric Spines.

BACKGROUND: Biomechanical properties of intact spinal motion segments are used to establish baseline values during in vitro studies evaluating spinal surgical techniques and implants. These properties are also used to validate computational models (ie, patient-specific finite element models) of human lumbar spine segments. Our laboratory has performed a large number of in vitro mechanical studies of lumbar spinal segments, using a consistent methodology. This provides extensive biomechanical data for a large number of intact motion segments, along with donor demographic variables, bone mineral density (BMD) measurements, and geometric properties. The objective of this study was to analyze how donor demographics, BMD, and geometric properties of cadaveric lumbar spine segments affect motion segment flexibility, including the range of motion (ROM), lax zone (LZ), and stiff zone (SZ), to help improve our understanding of spinal biomechanics. METHODS: A retrospective study examined the relationships between the biomechanical properties of 281 lumbar motion segments from 85 human cadaveric spines, donor demographic variables (age, sex, weight, height, and body mass index), and specimen measurements (vertebral body height, intervertebral disc height, and BMD). RESULTS: Statistical correlation and regression analyses showed that the flexibility of a lumbar motion segment is affected by lumbar level, donor age, sex, and weight as well as the intervertebral disc height, vertebral body height, and bone quality. Increased disc height was associated with decreased ROM (axial rotation), decreased LZ (flexion-extension and axial rotation), and increased SZ (flexion-extension and lateral bending) in the male group, but increased ROM (lateral bending) in the female group. Increased vertebral body height correlated with increased LZ (lateral bending) in the female group. Increased BMD correlated with decreased ROM overall. CONCLUSIONS: Biomechanical measurements from flexibility testing of cadaveric lumbar spine segments are significantly correlated with donor demographics and specimen measurements. Many of these correlations are sex-dependent.

Dimensional Characterization of the Human Cervical Interlaminar Space as a Guide for Safe Application of Minimally Invasive Dilators.

BACKGROUND: The risk of interlaminar passage of a dilator into the cervical spinal canal in minimally invasive approaches is currently unknown. Among the various anthropometric data reported in the literature, there is no report of the interlaminar dimensions in the cervical spine. OBJECTIVE: To report the cervical interlaminar dimensions in neutral, flexion, and extension. METHODS: A total of 8 spines were sectioned into cervical (C2-T1) segments. Digitized coordinate data defining the locations and movements of chosen anatomic points on the laminar edges at a given spinal level were used to compute the dimensions during a static neutral posture, flexion, and extension positions to mimic the positions during surgery. Interlaminar dimensions were averaged and categorized for each vertebral level and spinal posture. RESULTS: Based on the reported measurements, the smallest diameter dilator in commonly used dilator sets has the potential to traverse the interlaminar space at all levels in flexion. In a neutral posture, the average interlaminar distance at C2-3, C6-7, and C7-T1 was still greater than 2.0 mm, the smallest diameter of the initial dilator. The largest interlaminar distance was at C6-7 in flexion (7.68 ± 1.60 mm). CONCLUSION: Because dilators pass directly onto the cervical lamina without visualization of the midline structures, the interlaminar distances have increased relevance in the minimally invasive cervical approaches of foraminotomy and laminectomy. The data in this report demonstrate the theoretical risk of interlaminar passage with small diameter dilators in posterior minimally invasive approaches to the cervical spine.

Evaluation of abnormal styloid anatomy as a cause of internal jugular vein compression using a 3D-printed model: a laboratory investigation.

This study used a 3-dimensional (3D) craniocervical junction model of styloidogenic jugular venous compression (SJVC) syndrome to simulate and evaluate intracranial pressure (ICP) after internal jugular vein (IJV) compression by an elongated styloid process during axial rotation. The 3D-printed model created using data from an SJVC-syndrome patient included an articulating occipital-cervical junction, simplified arteriovenous system, gauge to measure simulated ICP, fixed obstruction simulating left-sided venous occlusion, and right-sided vascular tubing to simulate IJV compression. The model was rotated axially to its extreme right and left; maximum degree of motion and pressure were recorded for 3 cycles. Measurements were repeated after styloid resection in 25% increments. The extreme right rotation (11°) of the intact styloid condition yielded a mean pressure of 15.34 ±â€¯2.85 mmHg. After 25% styloid resection, extreme rotation (11°) yielded 13.96 ±â€¯2.88 mmHg. After 50%, extreme rotation increased to 16° yielding 17.41 ±â€¯3.52 mmHg; 11° rotation was 2.76 ±â€¯1.96 mmHg. After 75%, extreme rotation increased to 19° yielding -0.86 ±â€¯1.08 mmHg; 16° and 11° rotation yielded -0.69 ±â€¯1.19 and -0.86 ±â€¯1.08 mmHg, respectively. After 100%, extreme rotation to 19° yielded -1.21 ±â€¯0.60 mmHg; 16° and 11° rotation yielded -0.34 ±â€¯0.30 and 0.00 ±â€¯0.00 mmHg, respectively. Extreme left rotations (11°) yielded mean pressures of -0.17 ±â€¯0.00 (intact), -0.17 ±â€¯0.30 (25%), 2.24 ±â€¯0.79 (50%), 0.34 ±â€¯0.30 (75%), and 0.17 ±â€¯0.30 mmHg (100%). Simulated ICP increased proportionally to maximum ipsilateral axial rotation, and was highest after 50% styloid resection. Contralateral axial rotation did not increase pressure. IJV compression was relieved at 75% resection, suggesting that partial (75%) or complete styloidectomy is a potentially efficacious treatment for SJVC syndrome.

In Vitro Biomechanical Evaluation of a Novel, Minimally Invasive, Sacroiliac Joint Fixation Device.

BACKGROUND: Sacroiliac (SI) joint pathology may result in low-back pain, which causes substantial disability. Treatment failure with operative management of SI pain may be related to incomplete fusion of the joint and to fixation failure. The objective of this study was to evaluate the initial biomechanical stability of SI joint fixation with a novel implantable device in an in vitro human cadaveric model. METHODS: The right and left sides of 3 cadaveric L4-pelvis specimens were tested (1) intact, (2) destabilized, and (3) instrumented with an implantable SI joint fixation device using a simulated single-stance load condition. Right-leg and left-leg stance data were grouped together for a sample size of 6, and angular range of motion (ROM) was determined during application of flexion-extension, lateral bending, and axial rotation bending moments to a limit of 7.5 Nm. RESULTS: Following intact testing, destabilization by severing the posterior SI joint capsule and ligaments and the pubic symphysis reliably produced a significantly destabilized joint with the mean angular ROM more than doubling in flexion-extension and lateral bending and more than tripling in axial rotation (P ≤ .003) compared to the intact condition. Instrumentation with the SI screw fixation device significantly reduced mean joint ROM compared to the destabilized condition in all 3 anatomic planes tested (P < .001). When compared to the intact condition, the SI-instrumented condition significantly reduced lateral bending (P = .01) and had a similar ROM in flexion-extension (P = .14) and axial rotation (P = .85). CONCLUSIONS: Instrumentation with the SI screw fixation device significantly reduced mean joint ROM compared to the destabilized condition, with similar ROM in flexion-extension and axial rotation, and it significantly reduced ROM in lateral bending compared to that for the intact joint. The ROM values observed with the instrumented condition were comparable to levels of mobility considered favorable for spinal fusion.

The histone deacetylase inhibitor Entinostat enhances polymer-mediated transgene expression in cancer cell lines.

Eukaryotic cells maintain an immense amount of genetic information by tightly wrapping their DNA around positively charged histones. While this strategy allows human cells to maintain more than 25,000 genes, histone binding can also block gene expression. Consequently, cells express histone acetyl transferases (HATs) to acetylate histone lysines and release DNA for transcription. Conversely, histone deacetylases (HDACs) are employed for restoring the positive charge on the histones, thereby silencing gene expression by increasing histone-DNA binding. It has previously been shown that histones bind and silence viral DNA, while hyperacetylation of histones via HDAC inhibition restores viral gene expression. In this study, we demonstrate that treatment with Entinostat, an HDAC inhibitor, enhances transgene (luciferase) expression by up to 25-fold in human prostate and murine bladder cancer cell lines when used with cationic polymers for plasmid DNA delivery. Entinostat treatment altered cell cycle progression, resulting in a significant increase in the fraction of cells present in the G0/G1 phase at low micromolar concentrations. While this moderate G0/G1 arrest disappeared at higher concentrations, a modest increase in the fraction of apoptotic cells and a decrease in cell proliferation were observed, consistent with the known anticancer effects of the drug. DNase accessibility studies revealed no significant change in plasmid transcriptional availability with Entinostat treatment. However, quantitative PCR studies indicated that Entinostat treatment, at the optimal dose for enhancing transgene expression, led to an increase in the amount of plasmid present in the nucleus in two cancer cell lines. Taken together, our results show that Entinostat enhances polymer- mediated transgene expression and can be useful in applications related to transient protein expression in mammalian cells. Biotechnol. Bioeng. 2016;113: 1345-1356. © 2015 Wiley Periodicals, Inc.

A Phase I Double Blind, Placebo-Controlled, Randomized Study of the Safety and Immunogenicity of Electroporated HIV DNA with or without Interleukin 12 in Prime-Boost Combinations with an Ad35 HIV Vaccine in Healthy HIV-Seronegative African Adults.

BACKGROUND: Strategies to enhance the immunogenicity of DNA vaccines in humans include i) co-administration of molecular adjuvants, ii) intramuscular administration followed by in vivo electroporation (IM/EP) and/or iii) boosting with a different vaccine. Combining these strategies provided protection of macaques challenged with SIV; this clinical trial was designed to mimic the vaccine regimen in the SIV study. METHODS: Seventy five healthy, HIV-seronegative adults were enrolled into a phase 1, randomized, double-blind, placebo-controlled trial. Multi-antigenic HIV (HIVMAG) plasmid DNA (pDNA) vaccine alone or co-administered with pDNA encoding human Interleukin 12 (IL-12) (GENEVAX IL-12) given by IM/EP using the TriGrid Delivery System was tested in different prime-boost regimens with recombinant Ad35 HIV vaccine given IM. RESULTS: All local reactions but one were mild or moderate. Systemic reactions and unsolicited adverse events including laboratory abnormalities did not differ between vaccine and placebo recipients. No serious adverse events (SAEs) were reported. T cell and antibody response rates after HIVMAG (x3) prime-Ad35 (x1) boost were independent of IL-12, while the magnitude of interferon gamma (IFN-γ) ELISPOT responses was highest after HIVMAG (x3) without IL-12. The quality and phenotype of T cell responses shown by intracellular cytokine staining (ICS) were similar between groups. Inhibition of HIV replication by autologous T cells was demonstrated after HIVMAG (x3) prime and was boosted after Ad35. HIV specific antibodies were detected only after Ad35 boost, although there was a priming effect with 3 doses of HIVMAG with or without IL-12. No anti-IL-12 antibodies were detected. CONCLUSION: The vaccines were safe, well tolerated and moderately immunogenic. Repeated administration IM/EP was well accepted. An adjuvant effect of co-administered plasmid IL-12 was not detected. TRIAL REGISTRATION: ClinicalTrials.gov NCT01496989.

Long-term follow-up of study participants from prophylactic HIV vaccine clinical trials in Africa.

Long-term safety is critical for the development and later use of a vaccine to prevent HIV/AIDS. Likewise, the persistence of vaccine-induced antibodies and their impact on HIV testing must be established. IAVI has sponsored several Phase I and IIA HIV vaccine trials enrolling healthy, HIV-seronegative African volunteers. Plasmid DNA and viral vector based vaccines were tested. No vaccine-related serious adverse events were reported. After completion of vaccine trials conducted between 2001-2007, both vaccine and placebo recipients were offered enrolment into an observational long-term follow-up study (LTFU) to monitor potential late health effects and persistence of immune responses. At scheduled 6-monthly clinic visits, a health questionnaire was administered; clinical events were recorded and graded for severity. Blood was drawn for HIV testing and cellular immune assays. 287 volunteers were enrolled; total follow-up after last vaccination was 1463 person years (median: 5.2 years). Ninety-three (93)% of volunteers reported good health at their last LTFU visit. Infectious diseases and injuries accounted for almost 50% of the 175 reported clinical events, of which over 95% were mild or moderate in severity. There were 30 six pregnancies, six incident HIV infections and 14 volunteers reported cases of social harm. Persistence of immune responses was rare. No safety signal was identified. No potentially vaccine-related medical condition, no immune mediated disease, or malignancy was reported. HIV vaccines studied in these trials had a low potential of induction of persisting HIV antibodies.

A phase 2 study to evaluate the safety and immunogenicity of a recombinant HIV type 1 vaccine based on adeno-associated virus.

The recombinant vaccine, tgAAC09, based on an adeno-associated virus serotype 2 (AAV2) vector encoding HIV-1 subtype C Gag, protease, and part of reverse transcriptase, induced robust T cell and antibody responses in nonhuman primates. In a previous phase I study in 80 healthy HIV-seronegative European and Indian adults, the vaccine was generally safe, well tolerated, and modestly immunogenic when administered once at doses up to 3 x 10(11) DRP. This phase II double-blind, randomized, placebo-controlled trial tested two administrations and a higher dosage of tgAAC009. Ninety-one healthy HIV-seronegative adults from three African countries were given one of three dosage levels of tgAAC09 (3 x 10(10), 3 x 10(11), or 3 x 10(12) DRP) intramuscularly, either at a 6- or 12-month interval; follow-up was 18 months. Overall, 65% and 57% of vaccine recipients experienced local and systemic signs and symptoms, respectively, most being mild. Frequency and severity were not dose related and were similar to those in placebo recipients. No vaccine-related serious adverse events were reported. Overall, HIV-specific T cell responses were detected by IFN-gamma ELISPOT in 17/69 (25%) vaccine recipients with 38% (10/26) responders in the highest dosage group. The response rate improved significantly with boosting at 6, but not 12 months, in the 3 x 10(11) and 3 x 10(12) dosage groups only. Neutralizing antibody titers to the AAV2 did not alter the frequency of immune responses to HIV. Two doses of tgAAC09 were well tolerated at the dosage levels given. Fewer than half the recipients of the highest vaccine dosage, 3 x 10(12) DRP, had T cell responses to HIV.

Broad and potent neutralizing antibodies from an African donor reveal a new HIV-1 vaccine target.

Broadly neutralizing antibodies (bNAbs), which develop over time in some HIV-1-infected individuals, define critical epitopes for HIV vaccine design. Using a systematic approach, we have examined neutralization breadth in the sera of about 1800 HIV-1-infected individuals, primarily infected with non-clade B viruses, and have selected donors for monoclonal antibody (mAb) generation. We then used a high-throughput neutralization screen of antibody-containing culture supernatants from about 30,000 activated memory B cells from a clade A-infected African donor to isolate two potent mAbs that target a broadly neutralizing epitope. This epitope is preferentially expressed on trimeric Envelope protein and spans conserved regions of variable loops of the gp120 subunit. The results provide a framework for the design of new vaccine candidates for the elicitation of bNAb responses.

A phase 1 study to evaluate the safety and immunogenicity of a recombinant HIV type 1 subtype C adeno-associated virus vaccine.

A novel prophylactic AIDS vaccine candidate, consisting of single-stranded DNA for HIV-1 subtype C gag, protease, and part of reverse transcriptase genes, enclosed within a recombinant adeno-associated virus serotype-2 protein capsid (tgAAC09) induced T cell responses and antibodies in nonhuman primates. In this randomized, dose escalation phase I trial, HIV-uninfected healthy volunteers (50 in Europe, 30 in India) received a single intramuscular injection of tgAAC09 at 3 x 10(9) DNase resistant particles (DRP) (n = 16), 3 x 10(10) DRP (n = 23), 3 x 10(11) DRP (n = 25), or placebo (n = 16). Twenty-one participants in Europe received a second (boost) dose of 3 x 10(11) DRP tgAAC09 or placebo at least 24 weeks after the first injection. The vaccine was safe and well-tolerated after initial and boost vaccination. Local and systemic reactogenicity was experienced by 13-25% of participants and was not dose related. No vaccine-related serious adverse events were reported. Modest HIV-specific T cell responses were detected in 7/64 vaccinees (40-385 SFC/10(6) PBMC), with 16% (4/25) responders in the highest dose group. All responses were to Gag epitopes. tgAAC09 appears to be safe, well-tolerated, and modestly immunogenic. Further evaluation of higher doses of tgAAC09 and boost injections is ongoing in Africa.