Distinct gene expression by expanded clones of quiescent memory CD4+ T cells harboring intact latent HIV-1 proviruses.

Antiretroviral therapy controls, but does not cure, HIV-1 infection due to a reservoir of rare CD4+ T cells harboring latent proviruses. Little is known about the transcriptional program of latent cells. Here, we report a strategy to enrich clones of latent cells carrying intact, replication-competent HIV-1 proviruses from blood based on their expression of unique T cell receptors. Latent cell enrichment enabled single-cell transcriptomic analysis of 1,050 CD4+ T cells belonging to expanded clones harboring intact HIV-1 proviruses from 6 different individuals. The analysis reveals that most of these cells are T effector memory cells that are enriched for expression of HLA-DR, HLA-DP, CD74, CCL5, granzymes A and K, cystatin F, LYAR, and DUSP2. We conclude that expanded clones of latent cells carrying intact HIV-1 proviruses persist preferentially in a distinct CD4+ T cell population, opening possibilities for eradication.

Evolution of antibody immunity to SARS-CoV-2.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected 78 million individuals and is responsible for over 1.7 million deaths to date. Infection is associated with the development of variable levels of antibodies with neutralizing activity, which can protect against infection in animal models1,2. Antibody levels decrease with time, but, to our knowledge, the nature and quality of the memory B cells that would be required to produce antibodies upon reinfection has not been examined. Here we report on the humoral memory response in a cohort of 87 individuals assessed at 1.3 and 6.2 months after infection with SARS-CoV-2. We find that titres of IgM and IgG antibodies against the receptor-binding domain (RBD) of the spike protein of SARS-CoV-2 decrease significantly over this time period, with IgA being less affected. Concurrently, neutralizing activity in plasma decreases by fivefold in pseudotype virus assays. By contrast, the number of RBD-specific memory B cells remains unchanged at 6.2 months after infection. Memory B cells display clonal turnover after 6.2 months, and the antibodies that they express have greater somatic hypermutation, resistance to RBD mutations and increased potency, indicative of continued evolution of the humoral response. Immunofluorescence and PCR analyses of intestinal biopsies obtained from asymptomatic individuals at 4 months after the onset of coronavirus disease 2019 (COVID-19) revealed the persistence of SARS-CoV-2 nucleic acids and immunoreactivity in the small bowel of 7 out of 14 individuals. We conclude that the memory B cell response to SARS-CoV-2 evolves between 1.3 and 6.2 months after infection in a manner that is consistent with antigen persistence.

Enhanced SARS-CoV-2 neutralization by dimeric IgA.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus that causes coronavirus disease 2019 (COVID-19), primarily infects cells at mucosal surfaces. Serum neutralizing antibody responses are variable and generally low in individuals that suffer mild forms of COVID-19. Although potent immunoglobulin G (IgG) antibodies can neutralize the virus, less is known about secretory antibodies such as IgA that might affect the initial viral spread and transmissibility from the mucosa. Here, we characterize the IgA response to SARS-CoV-2 in a cohort of 149 convalescent individuals after diagnosis with COVID-19. IgA responses in plasma generally correlated with IgG responses. Furthermore, clones of IgM-, IgG-, and IgA-producing B cells were derived from common progenitor cells. Plasma IgA monomers specific to SARS-CoV-2 proteins were demonstrated to be twofold less potent than IgG equivalents. However, IgA dimers, the primary form of antibody in the nasopharynx, were, on average, 15 times more potent than IgA monomers against the same target. Thus, dimeric IgA responses may be particularly valuable for protection against SARS-CoV-2 and for vaccine efficacy.

Recommendations for measuring HIV reservoir size in cure-directed clinical trials.

Therapeutic strategies are being clinically tested either to eradicate latent HIV reservoirs or to achieve virologic control in the absence of antiretroviral therapy. Attaining this goal will require a consensus on how best to measure the numbers of persistently infected cells with the potential to cause viral rebound after antiretroviral-therapy cessation in assessing the results of cure-directed strategies in vivo. Current measurements assess various aspects of the HIV provirus and its functionality and produce divergent results. Here, we provide recommendations from the BEAT-HIV Martin Delaney Collaboratory on which viral measurements should be prioritized in HIV-cure-directed clinical trials.

Engineered extracellular matrix components do not alter the immunomodulatory properties of mesenchymal stromal cells in vitro.

Mesenchymal stromal cells (MSCs) have emerged as promising candidates for regenerative therapies, including tissue engineering. Recently it has been reported that engineered extracellular matrix (ECM) components support the differentiation of MSCs into osteocytes and chondrocytes, indicating that ECM components may represent attractive carriers for MSC transplants to repair damaged tissues. However, little is known about the impact of engineered ECM components on the immunosuppressive properties of MSCs, which may essentially contribute to the prevention of allogeneic MSC transplant rejection. In the present study, we explored the potential of fibronectin, fibrillar collagen I, tropocollagen and collagen I/heparin to influence the immunosuppressive capacities of MSCs. We found that these ECM components do not modulate the capability of MSCs to inhibit the proliferation of anti-CD3/anti-CD28 antibody-stimulated CD4⁺ and CD8⁺ T cells and of lymphocytes in a mixed lymphocyte reaction. In addition, the potential of MSCs to impair the production of immunostimulatory IL-12 and to improve the release of immunosuppressive IL-10 by 6-sulpho LacNAc⁺ (slan) dendritic cells (DCs), representing a pro-inflammatory subset of human blood DCs, was not altered by the ECM components. Furthermore, ECM components do not influence the ability of MSCs to inhibit the slanDC-induced proliferation of CD4⁺ T cells. In conclusion, the used engineered ECMs maintain important immunosuppressive properties of MSCs, which support their suitablility as carriers for MSC transplants in tissue engineering.

Posterior atlanto-axial arthrodesis for fixation of odontoid nonunions.

STUDY DESIGN: A retrospective case series. OBJECTIVE: To determine the clinical and radiographic long-term results after posterior atlanto-axial arthrodesis of odontoid nonunions. SUMMARY OF BACKGROUND DATA: Nonunion of odontoid fractures is a relatively common and dreaded complication after surgical and nonoperative treatment of these injuries. Although there might be a significant rate, which require surgical stabilization due to atlanto-axial instability, only few publications have covered this issue and presented reliable long-term results. METHODS: We retrospectively analyzed the clinical and radiographic records of 9 (4 women and 5 men) patients with an average age of 68 (42-78) years at the time of injury who had undergone posterior atlanto-axial arthrodesis for surgical treatment of odontoid nonunions between 1988 and 2004. For posterior atlanto-axial arthrodesis, we performed either C1-C2 transarticular screw fixation, or posterior wiring and bone grafting, or a combination of these 2 techniques. RESULTS: Eight patients achieved a satisfactory clinical outcome and returned to their preinjury activity level. The Smiley-Webster scale showed an overall functional outcome score of 2.2, which was 0.9 points superior to the outcome score before surgery. Neurologic deficits after operative treatment of the odontoid nonunion were evaluated in 2 patients. In all the other patients with primary neurologic deficits or delayed neurologic sequelae we saw a full recovery. Solid bony fusion of the cervical arthrodesis was achieved in all of the patients. Failures of reduction or fixation were noted in 2 patients, but no reoperations were necessary. CONCLUSION: In summary, we had a satisfactory outcome after surgical treatment of odontoid nonunions in patients with atlanto-axial instability and severe motion pain at the cervical spine. With a bony union rate of 100% and a noticeable improvement of clinical results and neurologic function, posterior atlanto-axial arthrodesis seems to be an appropriate option for nonunited odontoid fractures that require surgical stabilization.

Surgical treatment of dens fractures in elderly patients.

BACKGROUND: A dens fracture is the most common cervical fracture in elderly patients. The purposes of this study were to analyze the functional and radiographic results after surgical treatment of dens fractures in patients over sixty-five years of age and to compare the two methods that were used for operative treatment. METHODS: We reviewed the cases of fifty-six patients, with an average age of 71.4 years at the time of surgery, who had undergone surgical treatment of a dens fracture from 1988 to 2002. Thirty-seven fractures were stabilized with anterior screw fixation, and nineteen fractures had posterior cervical arthrodesis. RESULTS: Forty-five patients returned to their preinjury activity level and were satisfied with their treatment. Thirty-five patients had a full range of neck movement, and forty-seven patients were free of pain. Technical failures occurred in eight patients. The thirty-seven patients treated with anterior screw fixation had a good clinical outcome, with fracture-healing in thirty-three patients (89%) and technical failure in five patients (14%). All nineteen patients treated with posterior cervical arthrodesis had fracture union, with technical failure in three patients, but the functional results were worse than those after anterior screw fixation. With the inclusion of the six patients who had been excluded from the clinical and radiographic review, the overall morbidity rate was 16% (ten of sixty-two patients) and the overall mortality rate was 6% (four of sixty-two patients). CONCLUSIONS: A satisfactory outcome can be achieved with surgical treatment of a dens fracture in geriatric patients. It appears that anterior screw fixation may maintain better mobility of the cervical spine, but it appears to be associated with a higher rate of fracture nonunion and a greater potential for reoperation.

Anterior screw fixation of odontoid fractures comparing younger and elderly patients.

STUDY DESIGN: A retrospective, comparative study. SUMMARY OF BACKGROUND DATA AND OBJECTIVES: Anterior screw fixation has become an accomplished treatment option for the management of odontoid fractures. In younger patients, it has shown encouraging results with low complication rates; whereas in geriatric trauma victims, it remains the subject of controversy. The purpose of this study was to determine functional and radiographic results after anterior screw fixation of Type II odontoid fractures, with the particular interest to compare the outcome between younger and elderly patients. MATERIAL/METHODS: We reviewed clinical and radiographic records of 110 patients with an average age of 54 years at the time of surgery after anterior double screw fixation of their odontoid fractures between 1990 and 2004. To compare functional and radiographic results between nongeriatric and geriatric patients, they were divided by age into 2 groups: Group A included patients 65 years of age or younger and Group B contained patients older than 65 years. RESULTS: A total of 95 patients had returned to their preinjury activity level and were satisfied with their treatment. The Smiley-Webster scale showed an overall outcome score of 1.42 with similar results in both groups (Group A, 1.34; Group B, 1.50). Bony fusion was achieved in 102 patients, failures of reduction or fixation occurred in 12 patients. Comparing between the 2 groups, we had a nonunion rate of 4% in younger individuals and a significantly higher rate of 12% in geriatric patients. Reoperation due to nonunion or technical failures was necessary in 8 patients. CONCLUSION: We had encouraging results using anterior screw fixation for surgical treatment of odontoid fractures and favor this method as preferred management strategy for stabilization of these fractures. Comparing between age groups, we had similar results on cervical spine function. With regards to fracture healing as well as morbidity and mortality, younger patients had a superior outcome.

Thromboembolic complications after spinal surgery in trauma patients.

BACKGROUND: Deep venous thrombosis (DVT) and pulmonary embolism (PE) may be significant complications following spinal surgery. The incidence rate ranges from 0.5% to 2.5% in patients with symptomatic thromboembolic disease and up to 15% in patients with non-symptomatic thrombotic complications. We determined the incidence of symptomatic thromboembolism after spinal surgery in patients with postoperative systemic prophylaxis and investigated general and specific risk factors for development of this disease. PATIENTS AND METHODS: We analyzed the clinical records of 978 patients who had undergone surgery of the spine because of trauma and who had been admitted to our level-I trauma center between 1980 and 2004. Spinal procedures included anterior and/or posterior spinal fusion, video-assisted thoracoscopic fusion, and spinal decompression. Symptomatic thromboembolic disease was diagnosed when patients showed significant clinical signs or symptoms of DVT or PE. In cases of DVT, diagnosis was confirmed by duplex scan of the lower limbs; in cases of PE, diagnosis was confirmed by CT-scanning of the thorax or at post mortem. RESULTS: The incidence rate of symptomatic thromboembolic complications was 2.2% (n 22). 17 patients showed clinical signs of deep venous thrombosis, with 4 of them developing pulmonary embolism subsequently. The other 5 patients developed pulmonary embolism without prior clinical signs of deep venous thrombosis. 6 patients died because of thromboembolic disease. Thromboembolic complications were more frequent in older patients and among males, as well as in patients with regular tobacco consumption and obesity. Thromboembolic complications were also seen more frequently in patients with surgical procedures at the lumbar spine, in patients with anterior spinal fusion, and in those with motor deficits in the lower extremities. INTERPRETATION: We found a rather low rate of clinically significant thromboembolic complications after spinal surgery because of trauma, compared to the results reported in the literature. Level of spinal surgery, surgical approach, and motor deficits in the lower extremities were identified as specific risk factors for DVT or PE. Age, sex, obesity and regular smoking were identified as general risk factors.