Soluble PD-L1 shows no association to relapse and overall survival in early stage non-small cell lung cancer (NSCLC).

BACKGROUND: Cancer immune evasion is critical in non-small cell lung cancer (NSCLC) and has been targeted by immunotherapy. High soluble (s)PD-L1 is associated with reduced survival and treatment failure in advanced stages. Here we evaluated the effects of sPD-L1 on T cells, relapse free survival, and overall survival in early stage NSCLC. METHODS: In vitro T cell stimulation was performed in the presence of sPD-L1 to evaluate its immunomodulatory activity. Data from The Cancer Genome Atlas (TCGA) were investigated for PD-L1 splice variants and enzymes involved in proteolytic cleavage (i.e. ADAM10). Plasma from 74 NSCLC (stage IA-IIIB), as well as an additional 73 (control cohort) patients was collected prior to curative surgery. Thereafter sPD-L1 levels from an immunosorbent assay were correlated with patient outcome. RESULTS: In vitro sPD-L1 inhibited IFN-γ production and proliferation of T cells and induced a terminal effector CD4 T cell subtype expressing CD27. Data from the TCGA demonstrated that elevated mRNA levels of ADAM10 is a negative predictor of outcome in NSCLC patients. To investigate the clinical relevance of these in vitro and TCGA findings, we quantified sPD-L1 in the plasma of early-stage NSCLC patients. In the first cohort we found significantly higher sPD-L1 levels in relapsing NSCLC patients, with a multivariate analysis revealing high sPD-L1 (>1000 pg/mL) as an independent predictor of survival. However, these findings could not be validated in two independent control cohorts. DISCUSSION: Although in vitro and TCGA data support the suppressive effect of sPD-L1 we were unable to translate this in our clinical setting. These results may be due to the small patient number and their heterogeneity as well as the lack of a standardized sPD-L1 ELISA. Our inconclusive results regarding the value of sPD-L1 in early stage NSCLC warrant assay validation and further investigation in larger (neo-)adjuvant trials.

Hyperbaric Oxygen Therapy and Tissue Regeneration: A Literature Survey.

By addressing the mechanisms involved in transcription, signaling, stress reaction, apoptosis and cell-death, cellular structure and cell-to-cell contacts, adhesion, migration as well as inflammation; HBO upregulates processes involved in repair while mechanisms perpetuating tissue damage are downregulated. Many experimental and clinical studies, respectively, cover wound healing, regeneration of neural tissue, of bone and cartilage, muscle, and cardiac tissue as well as intestinal barrier function. Following acute injury or in chronic healing problems HBO modulates proteins or molecules involved in inflammation, apoptosis, cell growth, neuro- and angiogenesis, scaffolding, perfusion, vascularization, and stem-cell mobilization, initiating repair by a variety of mechanisms, some of them based on the modulation of micro-RNAs. HBO affects the oxidative stress response via nuclear factor erythroid 2-related factor 2 (Nrf2) or c-Jun N-terminal peptide and downregulates inflammation by the modulation of high-mobility group protein B1 (HMGB-1), toll-like receptor 4 and 2 (TLR-4, TLR-2), nuclear factor kappa-B (NFκB), hypoxia-inducible factor (HIF-1α) and nitric oxide (NO•). HBO enhances stem-cell homeostasis via Wnt glycoproteins and mammalian target of rapamycin (mTOR) and improves cell repair, growth, and differentiation via the two latter but also by modulation of extracellular-signal regulated kinases (ERK) and the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT) pathway. The HBO-induced downregulation of matrix metalloproteinases-2 and 9 (MMP-2/-9), rho-associated protein kinase (ROCK) and integrins improve healing by tissue remodeling. Interestingly, the action of HBO on single effector proteins or molecules may involve both up- or downregulation, respectively, depending on their initial level. This probably mirrors a generally stabilizing potential of HBO that tends to restore the physiological balance rather than enhancing or counteracting single mechanisms.

Cellular receptors. effect of anti-alloantiserum on the recognition of transplantation antigens.

The possibility that a rat alloantiserum DA anti-Lewis possesses similar recognition structures for Lewis transplantation antigens, as do DA immunocompetent cells, was investigated by raising an antiserum against this alloantiserum in (Lewis x DA)F(1) hosts. This antiserum, as well as one provoked by injecting DA lymphoid cells, was active against recognition structures for Lewis antigens of DA immunocompetent cells. The anti-(DA anti-Lewis) antiserum displayed the same degree of specificity as was found previously for anti-recognition structure sera prepared by injecting parental strain lymphoid cells into F(1) hosts. Since the activities of antisera raised against cell-bound receptors or against the antigen-binding region of an immunoglobulin were indistinguishable, it was concluded that the functional part of cell-associated receptors might be structurally similar to the variable portion of an immunoglobulin.

Cellular receptors: binding of radioactively labeled anti-alloantiserum.

RAT ALLOANTISERA OF THE FOLLOWING SPECIFICITIES WERE OBTAINED AFTER SINGLE SKIN GRAFT REJECTION: DA anti-BN, BN anti-DA, DA anti-Lewis, and Lewis anti-DA. Anti-alloantibodies were raised by injecting the first two alloantisera into (DA x BN)F(1) hosts and the last two alloantisera into (DA x Lewis)F(1) hosts. In this manner, four anti-alloantisera were raised: anti-(DA anti-BN), anti-(BN anti-DA), anti-(DA anti-Lewis), and anti-(Lewis anti-DA). From each anti-alloantiserum a gamma globulin fraction was prepared and trace labeled with (125)I. The labeled anti-alloantibodies could be shown to be fixed preferentially by red cells coated with the alloantibody used to induce them. They were also preferentially fixed by normal rat lymphoid cells presumed to carry recognition structures similar to alloantibody. For instance, anti-(DA anti-BN) was fixed by BN red cells coated with DA anti-BN and by normal DA lymphoid cells presumed to carry RS(BN). Conversely, anti-(BN anti-DA) was fixed by DA red cells coated with BN anti-DA and by normal BN lymphoid cells, carrying RS(DA). Absorption and inhibition experiments showed that alloantibodies and normal lymphoid cells compete for the same labeled anti-alloantibodies. We conclude that cellular receptors and circulating alloantibody are antigenically similar: They have the same aliotype.

Viral oncolysis: increased immunogenicity of host cell antigen associated with influenza virus.

A2G mice could be solidly immunized against the Ehrlich ascites tumor by single intraperitoneal injections of homogenized and lyophilized tumor cells which had been infected with oncolytic strains of influenza A virus. Similar homogenates from noninfected tumor cells were not immunogenic, even when mixed with egg-grown virus. The immunizing principle in viral oncolysates could not be separated from the oncolytic virus by differential centrifugation or adsorption to and elution from red cells. It could be inhibited by antibody raised in rabbits against the egg-grown oncolytic virus. This reaction showed serologic specificity. Thus, the immunogenicity of an oncolysate produced with the WSA strain of neurotropic influenza virus could be inhibited by rabbit anti-WSA, but not by rabbit antibody to the TUR strain of fowl plague virus. Conversely, the immunogenicity of an oncolysate prepared with the TUR strain could be inhibited by rabbit anti-TUR, but not by anti-WSA. When mice were preimmunized (primed) with egg-grown WSA virus, their antitumor response to a later injection of WSA oncolysate was of the anamnestic type. Priming with egg-grown influenza B virus had no such effect. It was concluded that the immunogenicity of certain host cell components was greatly increased by incorporation into the makeup of the oncolytic virus.

Genetically determined, interferon-dependent resistance to influenza virus in mice.

The genetically determined resistance towards orthomyxoviruses exhibited by mice homozygous (A2G) or heterozygous (A2G X A/J) for the gene Mx was abolished or greatly diminished by treatment with anti-interferon globulin (AIF). AIF induced increased susceptibility to challenge with hepatotropic, neurotropic, and pneumotropic strains of influenza A virus. Hepatotropic virus titers in blood and livers of AIF-treated, Mx-bearing mice were higher by a factor of 10(3)--10(6) than those in untreated mice of the same genotype, and were comparable to those in genetically susceptible (untreated or AIF-treated) mice. Peritoneal macrophages from Mx-bearing untreated mice were resistant to challenge with a macrophage-adapted strain of influenza A virus even in the presence of AIF. However, when macrophages were taken from resistant mice injected with AIF and also cultivated in the presence of AIF, they were as susceptible to the virus as macrophages taken from susceptible mice. We conclude that interferons is an important factor in resistance to orthomyxoviruses governed by the gene Mx.

Natural, genetically determined resistance toward influenza virus in hemopoietic mouse chimeras. Role of mononuclear phagocytes.

Radiation chimeras produced by crosswise transfers of bone-marrow cell among histocompatible mice susceptible, or genetically resistant, to lethal challenge by a number of myxoviruses were used to test whether macrophage resistance (as assessed in vitro) and resistance of the animal (as measured in vivo), both previously shown to be brought about by the gene Mx, were causally related. 49 chimeras were tested individually, both of resistance of their macrophages to in vitro challenge with M-TUR (a strain of avian influenza virus A/Turkey/England/63 adapted to grow in cultured mouse peritoneal macrophages), and for resistance of the animal in vivo upon challenge with pneumotropic, neurotropic, or hepatotropic influenza viruses. Cultivated Kupffer cells and peritoneal macrophages harvested from chimeric mice expressed the resistance phenotype of the bone-marrow donor irrespective of the host environment in which they had differentiated. However, susceptibility or resistance in vivo was according to the genotype of the host. Thus, inborn resistance of radiation chimeras was found to be independent of Mx-gene expression in cells of the hemopoietic system.

Inborn resistance of mice to myxoviruses: macrophages express phenotype in vitro.

A strain of avian influenza A virus was adapted to grow in mouse peritoneal macrophages in vitro. The adapted strain, called M-TUR, induced a marked cytopathic effect in macrophages from susceptible mice. Mice homozygous (A2G) or heterozygous (F1 hybrids between A2G and several susceptible strains) for the gene Mx, shown previously to induce a high level of resistance towards lethal challenge by a number of myxoviruses in vivo, yielded peritoneal macrophages which were not affected by M-TUR. Peritoneal macrophages could be classified as resistant or susceptible to M-TUR without sacrificing the cell donor. Backcrosses were arranged between (A2G X A/J)F1 and A/J mice. 64 backcross animals could be tested individually both for resistance of their macrophages in vitro after challenge with M-TUR, and for resistance of the whole animal in vivo after challenge with NWS (a neurotropic variant of human influenza A virus). Macrophages from 36 backcross mice were classified as susceptible, and all of these mice died after challenge. Macrophages from 28 mice were classified as resistant, and 26 mice survived challenge. We conclude that resistance of macrophages and resistance of the whole animal are two facets of the same phenomenon.

Virus-specific interferon action. Protection of newborn Mx carriers against lethal infection with influenza virus.

The efficacy of interferon in antiviral protection of newborn mice differing at the Mx locus was investigated. Adult mice bearing the allele Mx exhibit a high degree of specific resistance toward lethal challenge with influenza viruses. In contrast, newborn Mx carriers are virtually as susceptible to influenza viruses as newborn mice devoid of Mx. Resistance can be abrogated by treating adult animals with anti-interferon serum. Here, we provide direct evidence of a virus-specific effect of interferon in vivo: newborn mice carrying the resistance gene Mx could be protected against lethal influenza virus infection with doses of interferon that were not protective in the absence of Mx. The efficacy of interferon towards a picornavirus (encephalomyocarditis virus) and a rhabdovirus (vesicular stomatitis virus) was independent of Mx.

Cell-bound receptors for alloantigens on normal lymphocytes. II. Antialloantibody serum contains specific factors reacting with relevant immunocompetent T lymphocytes.

The rat popliteal node graft-vs.-host assay was shown to depend on the presence of parental T cells in the inoculum. Antialloantisera raised in F(1) hybrid rats against alloantibodies of one parent directed at transplantation antigens of the other parent displayed some or all of the following specific effects on parental T cells: They inhibited local GvH by purified T-cell suspensions; they blocked the capacity of GvH-reactive cells to adsorb onto fibroblast monolayers of the relevant genotype; together with complement, they killed GvH-reactive cells.

Cell-bound receptors for alloantigens on normal lymphocytes. I. Characterization of receptor-carrying cells by the use of antibodies to alloantibodies.

Antialloantibodies were prepared in F(1) hybrid rats by immunization with alloantibodies from one parent raised against antigens of the other parent. The Ig fraction of such antialloantibodies was iodinated and used to investigate the nature of idiotype-carrying normal lymphoid cells. Lymphoid cell populations of the proper genotype fixed radioactive antialloantibody in proportion to their B-cell content. Activated T-cell populations, when depleted of B cells, did not fix significant amounts of radioactivity. Idiotype-carrying cells were sensitive to rabbit antirat Ig serum lysis and to antialloantiserum lysis, but not to rabbit anti-T-serum lysis. Also, the normal idiotype-containing B cell could be shown to have the expected antigen-binding specificity of its receptor. This was shown by the use of fibroblast cell monolayers that adsorbed specifically those cells capable of fixing the proper antialloantibody.

Resection in stage I/II non-small cell lung cancer.

In spite of the developments in chemo- and radiotherapy, surgery remains the mainstay of curative treatment of early stage non-small cell lung cancer (NSCLC). In stage Ia/Ib (T1, T2, N0), NSCLC lobectomy offers the best chance for cure, yielding survival rates of between 58 and 76%. Since the extent of mediastinal lymph node dissection does not seem to play a major prognostic role in stage Ia, video-thoracoscopic lobectomy yields equally good results as the open approach. Due to the necessity for a small thoracotomy when harvesting the specimen and the time-consuming lymph-node dissection minimally invasive lobar resections have failed to become routinely used. Minor resections, though sometimes necessary from the functional point of view, have a lower curative potential. They yield the best results if applied in tumors measuring less than 2 cm. Stage II, characterized by involvement of the N1-position and/or a more central tumor growth, has a 5-year survival of 45-52% and requires treatment by lobectomy or pneumonectomy. Sleeve resection may obviate the need for pneumonectomy in central upper-lobe tumors. In interlobar N1, however, pneumonectomy is indicated from the oncological point of view, since even meticulous lymph-node dissection is unable to achieve tumor control in this situation.

Influence of route of gastric transposition on oxygen supply at cervical oesophagogastric anastomoses.

BACKGROUND: The microcirculation and oxygen supply at the oesophagogastric anastomosis are crucial factors that influence anastomotic healing after oesophagectomy. METHODS: Twenty-nine patients (mean age 61.7 years) underwent gastric transposition via an orthotopic (14) or retrosternal (15) route. Interstitial partial pressure of oxygen (PO2) of the stomach in the anastomotic region was measured during oesophagectomy and in the intensive care unit. Interstitial PO2 values were determined after ligation of the short gastric vessels, after ligation of the left gastric artery, after forming the conduit and after gastric transposition. Postoperative measurements were recorded during endotracheal intubation, while breathing oxygen by mask or through the nose, and while breathing air. RESULTS: Interstitial PO2 levels were significantly higher before ligation of the left gastric artery than after ligation (mean 76.1 (95 per cent confidence interval 54.9 to 103.1) versus 44.9 (24.6 to 77.1) mmHg; P = 0.001). Levels were also higher following orthotopic transposition compared with the retrosternal route (68.2 (44.0 to 118.8) versus 24.6 (10.7 to 39.4) mmHg; P = 0.001) and during each postoperative measurement period. No differences were found between the various oxygen supply systems. CONCLUSION: Oxygen supply at the anastomosis of the gastric conduit reaches higher levels after orthotopic than retrosternal gastric transposition.

How to chase a red herring and come up with a smallmouth bass.

The collaboration between Alick Isaacs and myself started in the summer of 1956. Our initial project was to show, by electron microscopy, that interference between inactivated influenza virus and live virus involved the transfer of material from the interfering virus to the host cell. This approach failed for technical reasons. However, in the course of this work it appeared that more interfering activity remained in the system than we were entitled to expect. One possible explanation was that a substance, not identical with the initial interfering virus, was being generated. Subsequent experiments, aimed at checking this hypothesis, led to the description of interferon.

Preoperative Glasgow Prognostic Score as additional independent prognostic parameter for patients with esophageal cancer after curative esophagectomy.

BACKGROUND: Inflammation accelerates tumor growth followed by reduced survival in patients with cancer. The aim of this study was to evaluate the prognostic relevance of preoperatively increased levels of C-reactive protein (CRP) and the corresponding Glasgow Prognostic Score (GPS) on patients with esophageal carcinoma undergoing curative esophagectomy. METHODS: The data of 174 operated esophageal cancer patients were evaluated retrospectively. Patient's demographic and clinico-pathological data, tumor specific data, preoperative plasma levels of CRP and albumin, the corresponding GPS, overall survival (OS) and progression free survival (PFS) were assessed. RESULTS: 103 (59.2%) had adenocarcinoma and 71 (40.8%) had squamous cell carcinoma. 71 patients (43%) had elevated CRP concentrations. 118 patients (71%) had GPS 0, 41 (25%) GPS 1 and 8 (4%) GPS 2. Mean GPS was 0.3 (0-2). 5-year OS was higher in patients with normal CRP than in those with increased CRP (68% vs. 39%; p = 0.007). 5-year OS in patients with GPS 0 and GPS 1 and 2 were 65% and 31% (p = 0.001). 5-year OS for the whole cohort was 56% (1 year: 83%, 3 years: 64%). Recurrence rate was 16.1% closely associated with GPS (p = 0.002). Median follow-up was 23 months (0-118 months). In multivariate analysis GPS, lymph node involvement, T stage and tumor histology were the independent prognostic parameters (p = 0.004, <0.001, 0.035, 0.010). CONCLUSIONS: Preoperatively increased GPS is significantly associated with reduced postoperative survival and tumor recurrence. The GPS as an independent prognosticator should be interpreted together with the TNM stage when the further postoperative treatment has to be scheduled.

Hyperbaric oxygenation in the treatment of life-threatening isobutyl nitrite-induced methemoglobinemia--a case report.

Methemoglobinemia usually results from exposure to oxidizing substances such as nitrates or nitrites. Iron within hemoglobin is oxidized from the ferrous (Fe2+) state to the ferric (Fe3+) state, resulting in the inability to transport oxygen and carbon dioxide. Clinically, this condition causes functional cyanosis. As methemoglobin levels increase, patients show evidence of cellular hypoxia in all tissues. Death usually occurs when methemoglobin fractions approach 70% of total hemoglobin. We describe the case of a 35-year-old female patient with severe life-threatening isobutyl nitrite-induced methemoglobinemia of 75% of total hemoglobin. Toluidine-blue was administered as first-line antidotal therapy immediately, followed by hyperbaric oxygenation. The patient recovered uneventfully and could be discharged 3 days later.

Host-cell antigen potentiated by incomplete growth cycle of influenza virus.

Lysis of Ehrlich ascites tumor cells in mice was induced with the Hong Kong influenza A-strain HKH virus not previously adapted to the tumor. Despite high pathogenicity of HKH, mice not genetically resistant to the lethal action of myxoviruses survived the actue phase of oncolysis. Virus infection of tumor cells resulted in high titers of hemagglutinin with low infectivity which indicated incomplete virus growth. Serial passages of HKH in Ehrlich ascites tumors failed. HKH oncolysates induced solid antitumor immunity in several mouse strains, including those fully susceptible to the virus. The immunizing power of HKH oncolysates could be abolished by mouse antibody against egg-grown HKH (H3, N2) but not by antiserum raised aganist TUR virus (Havl, Nav3).