PD-1-specific "Blocking" antibodies that deplete PD-1+ T cells present an inconvenient variable in preclinical immunotherapy experiments.

Therapeutic antibodies blocking PD-1-/PD-L1 interaction have achieved remarkable clinical success in cancer. In addition to blocking a target molecule, some isotypes of antibodies can activate complement, NK cells or phagocytes, resulting in death of the cell expressing the antibody's target. Human anti-PD-1 therapeutics use antibody isotypes designed to minimize such antibody-dependent lysis. In contrast, anti-PD-1 reagents used in mice are derived from multiple species, with different isotypes, and are not engineered to reduce target cell death: few studies analyze or discuss how antibody species and isotype may impact data interpretation. We demonstrate here that anti-PD-1 therapy to promote activation and proliferation of murine PD-1-expressing CD8 T cells sometimes led instead to a loss of antigen specific cells. This phenomenon was seen in two tumor models and a model of virus infection, and varied with the clone of anti-PD-1 antibody. Additionally, we compared competition among anti-PD-1 clones to find a combination that allows detection of PD-1-expressing cells despite the presence of blocking anti-PD1 antibodies in vivo. These data bring attention to the possibility of unintended target cell depletion with some commonly used anti-mouse PD-1 clones, and should provide a valuable resource for the design and interpretation of anti-PD-1 studies in mice.

Atividade antiviral e virucida de extratos hidroalcoólicos de própolis marrom, verde e de abelhas Jataí (Tetragonisca angustula) frente ao herpersvírus bovino tipo 1 (BoHV-1) e ao vírus da diarreia viral bovina (BVDV) / Antiviral and virucidal activity of hydroalcoholic extracts of propolis brown, green and jataí bees (Tetragonisca angustula) against Bovine Herpesvirus Type-1 (BoHV-1) and Bovine Viral Diarrhea Virus (BVDV)

Dentre as propriedades biológicas da própolis, a atividade antimicrobiana tem merecido destacada atenção. No presente trabalho, descreve-se a ação antiviral e virucida de três extratos hidroalcoólicos de própolis (marrom, verde e de abelhas jataí (Tetragonisca angustula), frente ao Herpesvírus Bovino tipo (BoHV-1) e ao Vírus da Diarreia Viral Bovina (BVDV). Os três extratos hidroalcoólicos foram obtidos de extração etanólica e são oriundos do sul do Brasil. A composição química dos extratos de própolis foi determinada pela cromatografia líquida de alta eficiência acoplada a espectrômetro de massas (UFLC-PDA-ESI-TOF/MS) que identificou e quantificou compostos como: ácido cafeico e ácido p-cumárico, ácido clorogênico, ácido ferúlico, além de flavonoides como a rutina. A toxicidade celular bem como a atividade antiviral dos extratos de própolis em monocamadas de células MDBK (Madin-Darby Bovine Kidney) foi avaliada através de observação microscópica e quantificada pelo teste de MTT (3-(4,5 dimetiltiazol-2yl)-2-5-difenil-2H tetrazolato de bromo). O extrato de própolis de abelhas jataí demonstrou ser menos citotóxico (1,57µg/mL), quando comparado aos extratos verde (0,78µg/mL) e marrom (0,39µg/mL). Quanto a atividade antiviral, a própolis verde demostrou maior eficácia em ambos os tratamentos celulares (pós e pré-exposição) frente ao BoHV-1 em relação aos outros extratos, ou seja, houve maior viabilidade celular quando comparada aos controles de células e vírus. Já a de jataí apresentou atividade frente aos dois vírus (BoHV-1 e BVDV) no método pré-infecção, enquanto a própolis marrom demonstrou ação apenas frente ao BoHV-1 também no método pré-infecção. Para determinação da atividade virucida foram utilizadas diferentes diluições dos vírus, bem como temperaturas e tempos distintos de incubação. A própolis verde a 37°C propiciou a maior redução no título viral (4,33log) em relação a marrom (log = 3,5log) e de jataí (log = 3,24log). No entanto, frente ao BVDV a própolis jataí apresentou os melhores resultados em ambas as temperaturas (22oC e 37oC). Portanto, os extratos avaliados apresentaram atividade antiviral e virucida frente ao BoHV-1 e BVDV, o que os torna alvo para o desenvolvimento de novos biofármacos como alternativa ao uso de antivirais comerciais em Medicina Veterinária.(AU)

Among the biological properties of propolis, the antimicrobial activity has received prominent attention. In this paper, we describe the antiviral and virucidal effect of three hydroalcoholic extracts of propolis (brown, green and jataí bees (Tetragonisca angustula), against bovine herpesvirus type-1 (BoHV-1) and bovine viral diarrhea Virus (BVDV). All hydroalcoholic extracts were obtained from ethanol extraction. The chemical composition of propolis extracts was determined by high-performance liquid chromatography coupled to mass spectrometer (UFLC-PDA-ESI-TOF/MS) to identify and quantify compounds such as caffeic acid and p-coumaric acid, chlorogenic acid, ferulic, and flavonoids such as rutin. Cell toxicity and antiviral activity of propolis extracts in monolayers of MDBK cells (Madin-Darby Bovine Kidney) were assessed by microscopic observation and quantified by the MTT assay (3- (4.5 dimethylthiazol-2yl) -2- 5-diphenyl-2H-tetrazolato bromine). Propolis extract from Jataí bees proved to be less cytotoxic (1.57mg / ml) when compared to green extracts (0.78mg / ml) and brown (0.39mg/mL). Regarding antiviral activity, propolis has shown greater efficacy in both cellular treatments (post and pre-exposure) against BoHV-1 when compared to other extracts, ie, there was increased cell viability compared to cell and virus controls. Extracts from Jataí showed activity against both viruses (BoHV-1 and BVDV) infection in the pre-test, whereas brown propolis demonstrated action only against the BoHV-1 in the pre-infection method. To determine the virucidal activity, it were used different dilutions of virus, as well as different temperatures and incubation times. The green propolis at 37°C led to a greater reduction in viral titer (4.33log) compared to brown (3.5log) and jataí (3.24log). Jataí propolis showed the best results in both temperatures (22oC and 37oC) when tested against BVDV. In summary, the evaluated extracts showed antiviral and virucidal activity against BoHV-1 and BVDV, and may be important targets for the development of new compounds as an alternative to commercial antivirals.(AU)

CRISPR/Cas9, a powerful tool to target human herpesviruses.

Over 90% of the adult population is infected with one or multiple herpesviruses. These viruses are characterized by their ability to establish latency, where the host is unable to clear the invader from infected cells resulting in a lifelong infection. Herpesviruses cause a wide variety of (recurrent) diseases such as cold sores, shingles, congenital defects and several malignancies. Although the productive phase of a herpesvirus infection can often be efficiently limited by nucleoside analogs, these drugs are ineffective during a latent herpesvirus infection and are therefore unable to clear herpesviruses from the human host. Advances in genome engineering using clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 facilitates virus research and may hold potential to treat or cure previously incurable herpesvirus infections by directly targeting these viruses within infected cells. Here, we review recent applications of the CRISPR/Cas9 system for herpesviral research and discuss the therapeutic potential of the system to treat, or even cure, productive and latent herpesviral infections.

Phototherapy in the treatment of cutaneous herpesvirus manifestations.

There is a considerable need for effective and safe treatment of cutaneous herpesvirus lesions. Current common approaches are limited to expensive or multidose oral pills. This systematic review of evidence-based approaches to phototherapy for the various manifestations of the herpesvirus discusses original publications of controlled clinical trials and case reports that were identified through searches in PubMed, MEDLINE, and Ovid. Interventions included photodynamic therapy (PDT), UV light, and near-infrared lasers. Nearly all studies (10 of 11) saw reduction of most or all lesions and extended time before reactivation of the virus. Side effects often were minimal to nonexistent, usually mild erythema at sites of phototreatment. Serious side effects included first-degree burns and linear IgA dermatosis, which were not common. Evidence from the reviewed literature indicates that short-term efficacy from treatment with phototherapy is the most likely outcome. However, long-term effects and follow-up of this treatment modality are lacking but appear promising. We recommend future studies to include more patients, determine the most effective type of phototherapy, and assess long-term follow-up. Furthermore, light-based therapies can be considered a reasonable alternative in situations that preclude traditional drug-based therapies.

Is cupping an effective treatment? An overview of systematic reviews.

Several systematic reviews (SRs) have assessed the effectiveness of cupping for a range of conditions. Our aim was to provide a critical evaluation and summary of these data. Electronic searches were conducted to locate all SRs concerning cupping for any condition. Data were extracted by two authors according to predefined criteria. Five SRs met our inclusion criteria, which related to the following conditions: pain conditions, stroke rehabilitation, hypertension, and herpes zoster. The numbers of studies included in each SR were small. Relatively clear evidence emerged only for one indication, that cupping may be effective for reducing pain. Based on evidence from the currently available SRs, the effectiveness of cupping has been demonstrated only as a treatment for pain, and even for this indication doubts remain.

Evidence-based interventional pain medicine according to clinical diagnoses. 17. Herpes zoster and post-herpetic neuralgia.

Herpes zoster infection is caused by a reactivation of the latent varicella zoster virus that causes chicken pox. It appears predominantly in older adults whose immunity for the virus has waned. The natural course of the disease is usually favorable, and the symptoms disappear spontaneously within a few weeks. Some patients, however, have prolonged pain: post-herpetic neuralgia. The diagnosis of acute zoster infection is made on the clinical signs including the appearance of rash. Post-herpetic neuralgia is described as sharp, burning, aching, or shooting constantly present in the dermatome that corresponds with the earlier rash. The objectives of treating herpes zoster are: (1) acute pain reduction; (2) promotion of recovery of epidermal defects and prevention of secondary infections; and (3) reduction or prevention of post-herpetic neuralgia. The objective of the treatment of post-herpetic neuralgia is primarily pain alleviation and improvement of the quality of life. Early treatment of the infection and the pain is believed to reduce the risk for post-herpetic neuralgia. This persistent pain syndrome is difficult to treat. Antiepileptic drugs and tricyclic antidepressants are the first choice. Interventional treatments, such as epidural injections of corticosteroids and local anesthetic drugs, have an effect on the acute pain but are of limited use in preventing post-herpetic neuralgia. When conservative treatment fails in providing satisfactory relief of post-herpetic neuralgia, a sympathetic block may be considered (2 C+); if this treatment provides unsatisfactory results, spinal cord stimulation may be considered, in a study context (2 C+).

Essential oils of peppermint, orange or lemongrass kill most strains of fungal and bacterial infections.

AIDS: A recent article claims that 10 essential oils (including lemongrass, eucalyptus, peppermint, and orange) were effective against 22 bacterial strains in laboratory studies. Lemongrass and orange oils completely inhibited 12 fungi, while peppermint oil eliminated 11 out of 12. A product that contains peppermint oil in an enteric-coated capsule is now available in health food stores. The capsule dissolves in the colon, where the peppermint oil should help kill off infections. HERP-EX, a blend of oils developed by Nature?s Grace (contact information is provided), is being used to successfully treat herpes 1 and 2. Patients are cautioned to research oils before using them.^ieng

Successful treatment of herpetic infections by autohemotherapy.

Herpes zoster (shingles) affects a significant number of individuals over age 50. To date, no satisfactory treatment has been available. The clinician author (JHO) witnessed a dramatic response of a shingles patient to autohemotherapy: the pain was completely relieved and lesions gone within 5 days with no recurrence of either. Treatment of other herpetic patients then began with autohemotherapy. Twenty-five patients with herpes were given an autologous blood transfer of 10 mL of blood from the antecubital vein into the gluteal bundle and followed for clinical signs. A 100% favorable response occurred in 20 patients who received autohemotherapy within 7 weeks of the onset of clinical signs and 1 other who received autohemotherapy at a 9-week interval. No untoward signs or symptoms of the treatment occurred. Autohemotherapy has been demonstrated to be effective in elimination of clinical sequelae in these cases of herpes infections and these results justify further rigorous clinical investigation.

The influence of age on transfer factor treatment of cellular immunodeficiency, chronic fatigue syndrome and/or chronic viral infections.

A group of 222 patients suffering from cellular immunodeficiency (CID), frequently combined with chronic fatigue syndrome (CFS) and/or chronic viral infections by Epstein-Barr virus (EBV) and/or cytomegalovirus (CMV), were immunologically investigated and treated with transfer factor (TF). The age range was 17-77 years. In order to elucidate the influence of aging on the course of the disease and on treatment, 3 subgroups were formed: 17-43 years, 44-53 years, and 54-77 years. Six injections of Immodin (commercial preparation of TF by SEVAC, Prague) were given in the course of 8 weeks. When active viral infection was present, IgG injections and vitamins were added. Immunological investigation was performed before the start of therapy, and subsequently according to need, but not later than after 3 months. The percentages of failures to improve clinical status of patients were in the individual subgroups, respectively: 10.6%, 11.5% and 28.9%. The influence of increasing age on the percentage of failures to normalize low numbers of T cells was very evident: 10.6%, 21.2% and 59.6%. In individuals uneffected by therapy, persistent absolute lymphocyte numbers below 1,200 cells were found in 23.1%, 54.5% and 89.3% in the oldest group. Statistical analysis by Pearson's Chi-square test, and the test for linear trend proved that the differences among the individual age groups were significant. Neither sex, nor other factors seemed to influence the results. The results of this pilot study show that age substantially influences the failure rate of CID treatment using TF. In older people, it is easier to improve the clinical condition than CID: this may be related to the diminished number of lymphocytes, however, a placebo effect cannot be totally excluded.

[So-called post-herpetic neuralgia and phantom limb pain: therapeutic concepts needing review]. / La cosidetta nevralgia posterpetica e l' arto fantasma doloroso: concetti terapeutici da rivedere.

After a brief review of the most recent findings on peripheral nerve nociception and deafferentation, the advisability of considering two so-called neuralgias - postherpetic neuralgia and phantom limb pain - in the more complex framework of deafferentation syndromes is examined together with the consequent need for a basic review of the therapeutic approaches pursued up to the present in the antalgic treatment of these two syndromes.

[Anterior and posterior medullary analgesic stimulation, using a percutaneous implantation technic]. / Stimulation analgésique médullaire antérieure et postérieure par technique d'implantation percutanée.

The technique of percutaneous implantation of a pain-relieving stimulator reduces the surgical procedure considerably. This advantage is, however, lessened by the absence of precision in placing epidural electrodes, and by the risk of their later displacement leading to inadequate stimulation. To reduce this disadvantage the authors suggest a technique that is a modification of the classical open procedure: it is particularly suitable for implantation at the cervico-dorsal junction. In 12 patients treated by percutaneous implantation, 5 received, from epidural electrodes, an anterior medullary electrical stimulation which caused analgesia without paraesthesiae in the painful ares. The results, however, do not justify a conclusion that anterior medullary stimulation is better than posterior column stimulation.

Genital herpes: Treatment with methylene blue and light exposure.

Fifty-six patients with genital herpes were treated by photodynamic inactivation of herpesvirus by methylene blue (36 patients), proflavine (101 or neutral red (10). The first 35 patients were randomly selected for treatment with one of the three dyes. All the others were initially treated with methylene blue. The disease was temporarily eradicated in about 70% of instances by exposure of the lesions to light plus topically applied methylene blue, neutral red or proflavine. Relapses, however, were not prevented or appreciably reduced.