Chemokine profiling of melanoma-macrophage crosstalk identifies CCL8 and CCL15 as prognostic factors in cutaneous melanoma.

During cancer evolution, tumor cells attract and dynamically interact with monocytes/macrophages. To find biomarkers of disease progression in human melanoma, we used unbiased RNA sequencing and secretome analyses of tumor-macrophage co-cultures. Pathway analysis of genes differentially modulated in human macrophages exposed to melanoma cells revealed a general upregulation of inflammatory hallmark gene sets, particularly chemokines. A selective group of chemokines, including CCL8, CCL15, and CCL20, was actively secreted upon melanoma-macrophage co-culture. Because we previously described the role of CCL20 in melanoma, we focused our study on CCL8 and CCL15 and confirmed that in vitro both chemokines contributed to melanoma survival, proliferation, and 3D invasion through CCR1 signaling. In vivo, both chemokines enhanced primary tumor growth, spontaneous lung metastasis, and circulating tumor cell survival and lung colonization in mouse xenograft models. Finally, we explored the clinical significance of CCL8 and CCL15 expression in human skin melanoma, screening a collection of 67 primary melanoma samples, using multicolor fluorescence and quantitative image analysis of chemokine-chemokine receptor content at the single-cell level. Primary skin melanomas displayed high CCR1 expression, but there was no difference in its level of expression between metastatic and nonmetastatic cases. By contrast, comparative analysis of these two clinically divergent groups showed a highly significant difference in the cancer cell content of CCL8 (p = 0.025) and CCL15 (p < 0.0001). Kaplan-Meier curves showed that a high content of CCL8 or CCL15 in cancer cells correlated with shorter disease-free and overall survival (log-rank test, p < 0.001). Our results highlight the role of CCL8 and CCL15, which are highly induced by melanoma-macrophage interactions in biologically aggressive primary melanomas and could be clinically applicable biomarkers for patient profiling. © 2024 The Pathological Society of Great Britain and Ireland.

ICAP-1 loss impairs CD8+ thymocyte development and leads to reduced marginal zone B cells in mice.

ICAP-1 regulates ß1-integrin activation and cell adhesion. Here, we used ICAP-1-null mice to study ICAP-1 potential involvement during immune cell development and function. Integrin α4ß1-dependent adhesion was comparable between ICAP-1-null and control thymocytes, but lack of ICAP-1 caused a defective single-positive (SP) CD8+ cell generation, thus, unveiling an ICAP-1 involvement in SP thymocyte development. ICAP-1 bears a nuclear localization signal and we found it displayed a strong nuclear distribution in thymocytes. Interestingly, there was a direct correlation between the lack of ICAP-1 and reduced levels in SP CD8+ thymocytes of Runx3, a transcription factor required for CD8+ thymocyte generation. In the spleen, ICAP-1 was found evenly distributed between cytoplasm and nuclear fractions, and ICAP-1-/- spleen T and B cells displayed upregulation of α4ß1-mediated adhesion, indicating that ICAP-1 negatively controls their attachment. Furthermore, CD3+ - and CD19+ -selected spleen cells from ICAP-1-null mice showed reduced proliferation in response to T- and B-cell stimuli, respectively. Finally, loss of ICAP-1 caused a remarkable decrease in marginal zone B- cell frequencies and a moderate increase in follicular B cells. Together, these data unravel an ICAP-1 involvement in the generation of SP CD8+ thymocytes and in the control of marginal zone B-cell numbers.

Inhibition of Orbivirus Replication by Aurintricarboxylic Acid.

Bluetongue virus (BTV) and African horse sickness virus (AHSV) are vector-borne viruses belonging to the Orbivirus genus, which are transmitted between hosts primarily by biting midges of the genus Culicoides. With recent BTV and AHSV outbreaks causing epidemics and important economy losses, there is a pressing need for efficacious drugs to treat and control the spread of these infections. The polyanionic aromatic compound aurintricarboxylic acid (ATA) has been shown to have a broad-spectrum antiviral activity. Here, we evaluated ATA as a potential antiviral compound against Orbivirus infections in both mammalian and insect cells. Notably, ATA was able to prevent the replication of BTV and AHSV in both cell types in a time- and concentration-dependent manner. In addition, we evaluated the effect of ATA in vivo using a mouse model of infection. ATA did not protect mice against a lethal challenge with BTV or AHSV, most probably due to the in vivo effect of ATA on immune system regulation. Overall, these results demonstrate that ATA has inhibitory activity against Orbivirus replication in vitro, but further in vivo analysis will be required before considering it as a potential therapy for future clinical evaluation.

Safety and Efficacy upon Infection in Sheep with Rift Valley Fever Virus ZH548-rA2, a Triple Mutant Rescued Virus.

The introduction of three single nucleotide mutations into the genome of the virulent RVFV ZH548 strain allows for the rescue of a fully attenuated virus in mice (ZH548-rA2). These mutations are located in the viral genes encoding the RdRp and the non-structural protein NSs. This paper shows the results obtained after the subcutaneous inoculation of ZH548-rA2 in adult sheep and the subsequent challenge with the parental virus (ZH548-rC1). Inoculation with the ZH548-rA2 virus caused no detectable clinical or pathological effect in sheep, whereas inoculation of the parental rC1 virus caused lesions compatible with viral infection characterised by the presence of scattered hepatic necrosis. Viral infection was confirmed via immunohistochemistry, with hepatocytes within the necrotic foci appearing as the main cells immunolabelled against viral antigen. Furthermore, the inoculation of sheep with the rA2 virus prevented the liver damage expected after rC1 virus inoculation, suggesting a protective efficacy in sheep which correlated with the induction of both humoral and cell-mediated immune responses.

Intratumoral BO-112 in combination with radiotherapy synergizes to achieve CD8 T-cell-mediated local tumor control.

BACKGROUND: Radioimmunotherapy combines irradiation of tumor lesions with immunotherapy to achieve local and abscopal control of cancer. Most immunotherapy agents are given systemically, but strategies for delivering immunotherapy locally are under clinical scrutiny to maximize efficacy and avoid toxicity. Local immunotherapy, by injecting various pathogen-associated molecular patterns, has shown efficacy both preclinically and clinically. BO-112 is a viral mimetic based on nanoplexed double-stranded RNA (poly I:C) which exerts immune-mediated antitumor effects in mice and humans on intratumoral delivery. BO-112 and focal irradiation were used to make the proof-of-concept for local immunotherapy plus radiation therapy combinations. METHODS: Murine transplantable tumor cell lines (TS/A, MC38 and B16-OVA) were used to show increased immunogenic features under irradiation, as well as in bilateral tumor models in which only one of the lesions was irradiated or/and injected with BO-112. Flow cytometry and multiplex tissue immunofluorescence were used to determine the effects on antitumor immunity. Depletions of immune cell populations and knockout mice for the IFNAR and BATF-3 genes were used to delineate the immune system requirements for efficacy. RESULTS: In cultures of TS/A breast cancer cells, the combination of irradiation and BO-112 showed more prominent features of immunogenic tumor cell death in terms of calreticulin exposure. Injection of BO-112 into the tumor lesion receiving radiation achieved excellent control of the treated tumor and modest delays in contralateral tumor progression. Local effects were associated with more prominent infiltrates of antitumor cytotoxic tumor lymphocytes (CTLs). Importantly, local irradiation plus BO-112 in one of the tumor lesions that enhanced the therapeutic effects of radiotherapy on distant irradiated lesions that were not injected with BO-112. Hence, this beneficial effect of local irradiation plus BO-112 on a tumor lesion enhanced the therapeutic response to radiotherapy on distant non-injected lesions. CONCLUSION: This study demonstrates that local BO-112 immunotherapy and focal irradiation may act in synergy to achieve local tumor control. Irradiation plus BO-112 in one of the tumor lesions enhanced the therapeutic effects on distant irradiated lesions that were not injected with BO-112, suggesting strategies to treat oligometastatic patients with lesions susceptible to radiotherapy and with at least one tumor accessible for repeated BO-112 intratumoral injections.

Activin A Sustains the Metastatic Phenotype of Tumor-Associated Macrophages and Is a Prognostic Marker in Human Cutaneous Melanoma.

Tumor cells attract and dynamically interact with monocytes/macrophages to subvert their differentiation into tumor-associated macrophages (TAMs), which mainly promote immune suppression and neoplastic progression, but the pathways and microenvironmental cues governing their protumoral deviation are not completely understood. To identify the molecular pathways responsible for TAM differentiation, we screened the biomarkers secreted during melanoma‒macrophage interactions using Quantibody microarrays and RNA sequencing of macrophages. We found that activin A, a member of the transforming GF family, plays an instrumental role in the cross-talk between melanoma cells and monocytes/macrophages, which results in the upregulation of distinct tumor-sustaining genes and the achievement of proinvasive and immunosuppressive functions of TAMs. Blockade of activin reduces the upregulation of part of these genes and prevents the acquisition of protumoral functions, facilitating human melanoma rejection by transferred human lymphocytes in a xenograft mouse model. Remarkably, screening of two independent cutaneous primary melanoma collections showed that activin A is enriched in TAMs and melanoma cells from patients with worse outcomes and constitutes a new and independent prognostic marker. Thus, we identify activin A as a key intermediary in the protumoral and immunosuppressive functions of TAMs, with significant potential as a disease biomarker as well as an immunotherapeutic target.

Status of novel cardiovascular risk factor and cardiovascular disease risk in an urban Cuban population--a pilot study.

Cardiovascular disease is the main cause of death in Cuba, yet the prevalence of novel risk factors is not known. To examine the prevalence of risk factors of traditional and novel cardiovascular diseases (CVDs) among an urban Cuban population, a cross-sectional pilot survey was undertaken in Havana city, Cuba. Ninety-seven adults aged 45-60 years registered to receive medical care at a policlinic. The prevalences of rates of CVD risk factors were: hypertension (> or =140/90 mmHg) (53.6%), hypercholesterolaemia (total cholesterol >5.2 mmol/L) (47.0%), low high-density lipoprotein (HDL)-cholesterol (<1.03 mmol/L) (64.3%); diabetes (self-reported) (24.6%); metabolic syndrome (ATP III criteria) (58.2%); overweight and obesity (body mass index > or = 25 kg/m2) (78.0%); current smoking (39.3%); elevated level of C-reactive protein (3 15 micromol/L) (11.1%). The total carotenoid status was independently associated with waist-circumference and risk of diabetes and metabolic syndrome. In this small unrepresentative sample of people aged 40-65 years from Havana city, there was a high prevalence of traditional and novel CVD risk factors. The total serum carotenoid status appeared to be associated with an increased prevalence of CVD risk factors.

The Macrophage Reprogramming Ability of Antifolates Reveals Soluble CD14 as a Potential Biomarker for Methotrexate Response in Rheumatoid Arthritis.

The identification of "trained immunity/tolerance" in myeloid cells has changed our perception of the performance of monocytes and macrophages during inflammatory and immune responses. Pemetrexed (PMX) and methotrexate (MTX) are blockers of the one-carbon metabolism (OCM) and commonly used therapeutic agents in cancer and rheumatoid arthritis (RA). We have previously showed that MTX promotes trained immunity in human macrophages. In the present manuscript, we have assessed the anti-inflammatory effects of PMX and MTX and found that OCM blockers alter the functional and gene expression profile of human macrophages and that OCM blockade reprograms macrophages towards a state of lipopolysaccharide (LPS) tolerance at the signaling and functional levels. Moreover, OCM blockade reduced macrophage LPS responsiveness by impairing the expression of membrane-bound and soluble CD14 (sCD14). The therapeutic relevance of these results was later confirmed in early RA patients, as MTX-responder RA patients exhibit lower sCD14 serum levels, with baseline sCD14 levels predicting MTX response. As a whole, our results demonstrate that OCM is a metabolic circuit that critically mediates the acquisition of innate immune tolerance and positions sCD14 as a valuable tool to predict MTX response in RA patients.

CCL20/TNF/VEGFA Cytokine Secretory Phenotype of Tumor-Associated Macrophages Is a Negative Prognostic Factor in Cutaneous Melanoma.

TAMs constitute a large fraction of infiltrating immune cells in melanoma tissues, but their significance for clinical outcomes remains unclear. We explored diverse TAM parameters in clinically relevant primary cutaneous melanoma samples, including density, location, size, and polarization marker expression; in addition, because cytokine production is a hallmark of macrophages function, we measured CCL20, TNF, and VEGFA intracellular cytokines by single-cell multiparametric confocal microscopy. The Kaplan-Meier method was used to analyze correlation with melanoma-specific disease-free survival and overall survival. No significant correlations with clinical parameters were observed for TAM density, morphology, or location. Significantly, higher contents of the intracellular cytokines CCL20, TNF, and VEGFA were quantified in TAMs infiltrating metastasizing compared to non-metastasizing skin primary melanomas (p < 0.001). To mechanistically explore cytokine up-regulation, we performed in vitro studies with melanoma-conditioned macrophages, using RNA-seq to explore involved pathways and specific inhibitors. We show that p53 and NF-κB coregulate CCL20, TNF, and VEGFA in melanoma-conditioned macrophages. These results delineate a clinically relevant pro-oncogenic cytokine profile of TAMs with prognostic significance in primary melanomas and point to the combined therapeutic targeting of NF-kB/p53 pathways to control the deviation of TAMs in melanoma.

Sexual behaviour, history of sexually transmitted diseases, and the risk of prostate cancer: a case-control study in Cuba.

BACKGROUND: The relationship between the risk of prostate cancer and sexual activity and history of sexually transmitted diseases was investigated in a case-control study conducted in Cuba aimed at assessing the effect of lifestyle and environmental factors, as well as hormonal and genetic factors, on the occurrence of this disease. METHODS: During the period 1998-2000, all men up to 84 yr old with newly diagnosed, cytologically and/or histologically confirmed prostatic cancer who were resident in Havana City were identified in nine major hospitals in the area. Controls were resident in the same city, frequency-matched by age (+/-5 years) and hospital. The study included 273 cases and 254 controls. Information was obtained through an interview. RESULTS: The risk of prostate cancer was increased among men with a history of venereal disease (odds ratio = 1.7, 95% CI = 1.1-2.5). A higher frequency of cases reported having had sex with prostitutes, although the estimate of relative risk did not reach statistical significance. Similarly, a nonsignificant positive association was found with the number of female sexual partners. A significant increased risk was observed in subjects who reported having sexual intercourse more than 7 times per week compared with those who reported a weekly frequency of 3 times or fewer (odds ratio = 2.1, 95% CI = 1.2-3.7). Moreover, a significant trend was demonstrated. CONCLUSIONS: The study supports the hypothesis that an infectious factor related to sexual behaviour could be involved in the occurrence of prostate cancer. A role for hormonal factors related to sexual activity cannot be ruled out.

[Ethical questions in the Spanish journal "Medicina Paliativa": 1994 - 2013]. / Cuestiones éticas en la revista medicina paliativa: 1994 - 2013.

OBJECTIVE: "Medicina Paliativa" is the official journal of the Spanish Society of Palliative Care ("Sociedad Española de Cuidados Paliativos"; SECPAL) and it reflects the interests, also on bioethics, of the professionals caring terminal people. We want to know what the bioethical questions they discuss and their approaches are. METHODS: From all the published articles in "Medicina Paliativa" from 1994 to 2013 we selected those referred to bioethics topics. We analysed: number of publications, author, subtype of article, year of publication, topic, philosophic approach, and the presence of answers and discussion. Qualitative topics were reviewed and agreed by at least two authors. RESULTS: There were 60 (9%) publications with bioethics profile from a total of 672 analyzed articles. A majority were signed by only one author. 31 (51%) were published as letter. The most relevant topics were: euthanasia, dignity, proportionality of treatment and care, sedation, principles of bioethics, and information. The orientation of these papers was, in a majority, according to principles of palliative care. Most bioethics topics have been answered and even answered back. CONCLUSION: Bioethics is an interesting topic in palliative care. The arguments usually fit the principles of palliative care.

Cuestiones éticas en la revista medicina paliativa: 1994 - 2013 / Ethical questions in the spanish journal "medicina paliativa": 1994 - 2013

Objetivo: La revista Medicina Paliativa, publicación oficial de la Sociedad Española de Cuidados Paliativos (SECPAL), refleja las inquietudes, también bioéticas, de los profesionales que atienden enfermos en situación terminal. Pretendemos conocer las cuestiones y los planteamientos bioéticos que debaten estos profesionales. Material y método: De los artículos publicados en Medicina Paliativa de 1994 a 2013 se recogen los que se centran en cuestiones bioéticas. Se analiza: número de trabajos, autores, tipo de artículo, año de publicación, tema, orientación y réplicas y/o debate. Las cuestiones de perfil cualitativo se han consensuado entre los revisores. Resultados: Se recogen 60 trabajos de perfil bioético, que suponen el 9% de los 672 analizados. La mayor parte tienen un solo autor. 31 (51%) se han publicado en formato de carta. Los temas más relevantes han sido: eutanasia, dignidad, proporcionalidad de las medidas, sedación, principios de bioética e información. La orientación de los trabajos se adapta en la mayoría de los casos a los principios de los Cuidados Paliativos. Las cuestiones bioéticas se han prestado al debate con réplicas y contrarréplicas. Conclusión: La bioética es un área de interés y debate en Cuidados Paliativos. Los argumentos se adaptan a los principios de los Cuidados Paliativos

Objective: "Medicina Paliativa" is the official journal of the Spanish Society of Palliative Care ("Sociedad Española de Cuidados Paliativos"; SECPAL) and it reflects the interests, also on bioethics, of the pro-fessionals caring terminal people. We want to know what the bioethical questions they discuss and their approaches are. Methods: From all the published articles in "Medicina Paliativa" from 1994 to 2013 we selected those referred to bioethics topics. We analysed: number of publications, author, subtype of article, year of publication, topic, philosophic approach, and the presence of answers and discussion. Qualitative topics were reviewed and agreed by at least two authors. Results: There were 60 (9%) publications with bioethics profile from a total of 672 analyzed articles. A majority were signed by only one author. 31 (51%) were published as letter. The most relevant topics were: euthanasia, dignity, proportionality of treatment and care, sedation, principles of bioethics, and information. The orientation of these papers was, in a majority, according to principles of palliative care. Most bioethics topics have been answered and even answered back. Conclusion: Bioethics is an interesting topic in palliative care. The arguments usually fit the principles of palliative care

Sexual behaviour, history of sexually transmitted diseases, and the risk of prostate cancer: a case–control study in Cuba

Background The relationship between the risk of prostate cancer and sexual activity and history of sexually transmitted diseases was investigated in a case–control study conducted in Cuba aimed at assessing the effect of lifestyle and environmental factors, as well as hormonal and genetic factors, on the occurrence of this disease. Methods During the period 1998–2000, all men up to 84 yr old with newly diagnosed, cytologically and/or histologically confirmed prostatic cancer who were resident in Havana City were identified in nine major hospitals in the area. Controls were resident in the same city, frequency-matched by age (±5 years) and hospital. The study included 273 cases and 254 controls. Information was obtained through an interview. Results The risk of prostate cancer was increased among men with a history of venereal disease (odds ratio = 1.7, 95 percent CI = 1.1–2.5). A higher frequency of cases reported having had sex with prostitutes, although the estimate of relative risk did not reach statistical significance. Similarly, a nonsignificant positive association was found with the number of female sexual partners. A significant increased risk was observed in subjects who reported having sexual intercourse more than 7 times per week compared with those who reported a weekly frequency of 3 times or fewer (odds ratio = 2.1, 95 percent CI = 1.2–3.7). Moreover, a significant trend was demonstrated. Conclusions The study supports the hypothesis that an infectious factor related to sexual behaviour could be involved in the occurrence of prostate cancer. A role for hormonal factors related to sexual activity cannot be ruled out(AU)

Antecedentes La relación entre el riesgo de cáncer de próstata y la actividad sexual y la historia de las enfermedades de transmisión sexual se ha investigado en un estudio caso-control realizado en Cuba, destinado a evaluar el efecto de estilo de vida y factores ambientales, así como los factores genéticos y hormonales, en el aparición de esta enfermedad. Métodos Durante el período 1998-2000, todos los hombres hasta 84 años de edad, con diagnóstico reciente, citológicamente y / o histológico de cáncer de próstata que son residentes en Ciudad de La Habana fueron identificados en nueve grandes hospitales de la zona. Los controles fueron residentes en la misma ciudad, la frecuencia con ajuste por edad (± 5 años) y el hospital. El estudio incluyó 273 casos y 254 controles. La información se obtuvo a través de una entrevista. Resultados El riesgo de cáncer de próstata se incrementó entre los hombres con un historial de enfermedades venéreas (odds ratio = 1,7, IC 95 por ciento = 1.1-2.5). Una mayor frecuencia de casos notificados de haber tenido relaciones sexuales con prostitutas, aunque la estimación del riesgo relativo no alcanzó significación estadística. Del mismo modo, uno no significativo se encontró asociación positiva con el número de parejas sexuales. Un importante aumento del riesgo se observó en los sujetos que reportaron tener relaciones sexuales más de 7 veces por semana en comparación con aquellos que informaron de una frecuencia semanal de 3 veces o menos (odds-ratio = 2,1, IC 95 por ciento = 1.2-3.7). Por otra parte, una tendencia importante fue demostrado. Conclusiones El estudio apoya la hipótesis de que un factor infecciosas relacionadas con el comportamiento sexual podrían estar implicados en la aparición del cáncer de próstata. El papel de los factores hormonales relacionados con la actividad sexual no se puede descartar

Estudio de casos y controles sobre factores de riesgo de cáncer de próstata / A case-control study on prostate cancer risks

Para determinar el papel de algunos factores genéticos, hormonales, de estilo de vida y ambientales en el riesgo de desarrollar cáncer de próstata clínicamente manifiesto en La Habana, se estudiaron los casos (n = 273) diagnosticados durante 1998-2000, con verificación histológica o citológica de cáncer de próstata, residentes en Ciudad de La Habana y menores de 85 años. Los controles (n = 254) fueron seleccionados pareados por edad, del mismo hospital del caso. La información se obtuvo mediante una entrevista. La estimación del riesgo se obtuvo a través de una regresión logística condicional. No se observó asociación estadísticamente significativa entre el color de la piel y el riesgo de cáncer de próstata (OR = 1,30, IC 95(por ciento): 0,92-1,84) ni con el hábito de fumar (OR = 0,82, IC 95(por ciento): 0,58-1,16). Se observó una asociación positiva del riesgo de cáncer de próstata con las enfermedades venéreas (p = 0,01), así como con la edad de aparición de estas enfermedades (p = 0,06). No se encontraron diferencias en cuanto a la edad de inicio de las relaciones sexuales (p = 0,111) ni en el número de compañeras sexuales (p = 0,48). Se observó una asociación significativa entre el riesgo de padecer cáncer de próstata y el haber padecido de alguna enfermedad venérea, el no realizar ejercicio físico entre los 45 y los 50 años y la frecuencia de relaciones sexuales por encima de 10 veces a la semana

Estudio de casos y controles sobre factores de riesgo de cáncer de próstata / A case-control study on prostate cancer risks

Para determinar el papel de algunos factores genéticos, hormonales, de estilo de vida y ambientales en el riesgo de desarrollar cáncer de próstata clínicamente manifiesto en La Habana, se estudiaron los casos (n = 273) diagnosticados durante 1998-2000, con verificación histológica o citológica de cáncer de próstata, residentes en Ciudad de La Habana y menores de 85 años. Los controles (n = 254) fueron seleccionados pareados por edad, del mismo hospital del caso. La información se obtuvo mediante una entrevista. La estimación del riesgo se obtuvo a través de una regresión logística condicional. No se observó asociación estadísticamente significativa entre el color de la piel y el riesgo de cáncer de próstata (OR = 1,30, IC 95(por ciento): 0,92-1,84) ni con el hábito de fumar (OR = 0,82, IC 95(por ciento): 0,58-1,16). Se observó una asociación positiva del riesgo de cáncer de próstata con las enfermedades venéreas (p = 0,01), así como con la edad de aparición de estas enfermedades (p = 0,06). No se encontraron diferencias en cuanto a la edad de inicio de las relaciones sexuales (p = 0,111) ni en el número de compañeras sexuales (p = 0,48). Se observó una asociación significativa entre el riesgo de padecer cáncer de próstata y el haber padecido de alguna enfermedad venérea, el no realizar ejercicio físico entre los 45 y los 50 años y la frecuencia de relaciones sexuales por encima de 10 veces a la semana(AU)