Neutralization-guided design of HIV-1 envelope trimers with high affinity for the unmutated common ancestor of CH235 lineage CD4bs broadly neutralizing antibodies

The CD4 binding site (CD4bs) of the HIV-1 envelope glycoprotein is susceptible to multiple lineages of broadly neutralizing antibodies (bnAbs) that are attractive to elicit with vaccines. The CH235 lineage (VH1-46) of CD4bs bnAbs is particularly attractive because the most mature members neutralize 90% of circulating strains, do not possess long HCDR3 regions, and do not contain insertions and deletions that may be difficult to induce. We used virus neutralization to measure the interaction of CH235 unmutated common ancestor (CH235 UCA) with functional Env trimers on infectious virions to guide immunogen design for this bnAb lineage. Two Env mutations were identified, one in loop D (N279K) and another in V5 (G458Y), that acted synergistically to render autologous CH505 transmitted/founder virus susceptible to neutralization by CH235 UCA. Man5-enriched N-glycans provided additional synergy for neutralization. CH235 UCA bound with nanomolar affinity to corresponding soluble native-like Env trimers as candidate immunogens. A cryo-EM structure of CH235 UCA bound to Man5-enriched CH505.N279K.G458Y.SOSIP.664 revealed interactions of the antibody light chain complementarity determining region 3 (CDR L3) with the engineered Env loops D and V5. These results demonstrate that virus neutralization can directly inform vaccine design and suggest a germline targeting and reverse engineering strategy to initiate and mature the CH235 bnAb lineage

Relatório de estágio na Câmara Municipal de Ourém

O presente relatório enquadra-se no âmbito do trabalho final do curso Mestrado em Reabilitação Urbana, do Instituto Politécnico de Tomar. O estágio decorreu na Câmara Municipal de Ourém e teve como objeto, participar na coordenação com a Divisão Obras Municipais, Divisão da Ação Cultural e Divisão de Educação e Assuntos Sociais em intervenções a nível da Reabilitação Urbana. O estágio numa Câmara Municipal foi a opção que pareceu mais enriquecedora para conclusão do Mestrado em Reabilitação Urbana e para o futuro da minha vida profissional. Esta opção de estágio deve-se ao desejo de poder aplicar alguns dos conhecimentos obtidos ao longo do Mestrado, de conhecer a realidade do trabalho em obra e inserido numa equipa profissional, fiscalizadora, lidando com as responsabilidades inerentes à profissão. O relatório aborda quatro casos de estudo, respeitante às reabilitações, 1ª fase de intervenção na capela de São Sebastião em Atouguia, 2ª fase da reabilitação de zona de lazer no Agroal, freguesia de Formigais, 1ª fase de intervenção da capela da Perucha e reforço do pontão de Caxarias. Acompanhamento de pequenas reabilitações, reclamadas por juntas de freguesia e munícipes. O relatório procura descrever os procedimentos adotados no acompanhamento das obras de reabilitação e para fácil entendimento as descrições são acompanhadas por fotografias

Activity of HIV Entry And Fusion Inhibitors Expressed By The Human Vaginal Colonizing Probiotic Lactobacillus Reuteri RC-14

Novel therapeutic approaches are needed to combat the rapid increase in HIV sexual transmission in women. The probiotic organism Lactobacillus reuteri RC-14 which safely colonizes the human vagina and prevents microbial infections, has been genetically modified to produce anti-HIV proteins which were capable of blocking the three main steps of HIV entry into human peripheral blood mononuclear cells. The HIV entry or fusion inhibitors were fused to the native expression and secretion signals of BspA, Mlp or Sep in L. reuteri RC-14 and the expression cassettes were stably inserted into the chromosome. L. reuteri RC-14 expressed the HIV inhibitors in cell wall-associated and secreted forms. L. reuteri RC-14 expressing CD4D1D2-antibody-like fusion proteins were able to bind single or dual tropic coreceptor-using HIV-1 primary isolates. This is the first study to show that a well-documented and proven human vaginal probiotic strain can express potent functional viral inhibitors, which may potentially lower the sexual transmission of HIV. © 2006 The Authors; Journal compilation © 2006 Blackwell Publishing Ltd

D614G Spike mutation increases SARS CoV-2 susceptibility to neutralization

The SARS-CoV-2 Spike protein acquired a D614G mutation early in the pandemic that confers greater infectivity and is now the globally dominant form. To determine whether D614G might also mediate neutralization-escape that could compromise vaccine efficacy, sera from Spike-immunized mice, nonhuman primates and humans were evaluated for neutralization of pseudoviruses bearing either D614 or G614 spike. In all cases, the G614 pseudovirus was moderately more susceptible to neutralization. The G614 pseudovirus also was more susceptible to neutralization by receptor binding domain (RBD) monoclonal antibodies and convalescent sera from people infected with either form of the virus. Negative stain electron microscopy revealed a higher percentage of the 1-RBD “up” conformation in the G614 spike, suggesting increased epitope exposure as a mechanism of enhanced vulnerability to neutralization. Based on these findings, the D614G mutation is not expected to be an obstacle for current vaccine development