Expression of CD11c Is Associated with Unconventional Activated T Cell Subsets with High Migratory Potential

Ajudes rebudes: Marie Curie Career Integration Grant; Dexeus Foundation for Women's Health Research; i Contratos Ramón y CajalCD11c is an α integrin classically employed to define myeloid dendritic cells. Although there is little information about CD11c expression on human T cells, mouse models have shown an association of CD11c expression with functionally relevant T cell subsets. In the context of genital tract infection, we have previously observed increased expression of CD11c in circulating T cells from mice and women. Microarray analyses of activated effector T cells expressing CD11c derived from naïve mice demonstrated enrichment for natural killer (NK) associated genes. Here we find that murine CD11c+ T cells analyzed by flow cytometry display markers associated with non-conventional T cell subsets, including γδ T cells and invariant natural killer T (iNKT) cells. However, in women, only γδ T cells and CD8+ T cells were enriched within the CD11c fraction of blood and cervical tissue. These CD11c+ cells were highly activated and had greater interferon (IFN)-γ secretory capacity than CD11c- T cells. Furthermore, circulating CD11c+ T cells were associated with the expression of multiple adhesion molecules in women, suggesting that these cells have high tissue homing potential. These data suggest that CD11c expression distinguishes a population of circulating T cells during bacterial infection with innate capacity and mucosal homing potential

Adhesion Molecules Associated with Female Genital Tract Infection

Altres ajuts: Marie Curie Career Integration Grant i una beca Fundació Dexeus Salut de la DonaEfforts to develop vaccines that can elicit mucosal immune responses in the female genital tract against sexually transmitted infections have been hampered by an inability to measure immune responses in these tissues. The differential expression of adhesion molecules is known to confer site-dependent homing of circulating effector T cells to mucosal tissues. Specific homing molecules have been defined that can be measured in blood as surrogate markers of local immunity (e.g. α4β7 for gut). Here we analyzed the expression pattern of adhesion molecules by circulating effector T cells following mucosal infection of the female genital tract in mice and during a symptomatic episode of vaginosis in women. While CCR2, CCR5, CXCR6 and CD11c were preferentially expressed in a mouse model of Chlamydia infection, only CCR5 and CD11c were clearly expressed by effector T cells during bacterial vaginosis in women. Other homing molecules previously suggested as required for homing to the genital mucosa such as α4β1 and α4β7 were also differentially expressed in these patients. However, CD11c expression, an integrin chain rarely analyzed in the context of T cell immunity, was the most consistently elevated in all activated effector CD8+ T cell subsets analyzed. This molecule was also induced after systemic infection in mice, suggesting that CD11c is not exclusive of genital tract infection. Still, its increase in response to genital tract disorders may represent a novel surrogate marker of mucosal immunity in women, and warrants further exploration for diagnostic and therapeutic purposes

Adhesion molecules and effector T cells subsets associated with female genital tract infection

Programa de Doctorat en Biomedicina[eng] Common sexually transmitted infections like gonorrhea, Chlamydia, syphilis, and trichomoniasis are the leading cause of morbidity in developing countries. Adolescents and young women have the highest rates of STIs in developing countries. Pre-existing female genital tract infections affect the development and pathogenesis of other sexually transmitted infections, leading to long-term consequences such as pelvic inflammatory disease, cancer, and infertility. Efforts to develop vaccines against sexually transmitted infections have been hindered by the inability to measure immune responses in the genital tract. Effective vaccines should generate localized immune responses at potential exposure sites to provide faster and more robust infection control. Assays rely on blood samples to provide a complete picture of cellular traffic between lymph nodes and the mucosa, especially soon after infection. Studying and characterizing the homing profile of T cells becomes crucial to understanding mucosal immune responses against infections, and new methods to measure and manipulate immune responses are needed to develop effective STI vaccines. In this study, we examined how adhesion molecules are expressed by circulating effector T cells following mucosal infection of the female genital tract in mice and during symptomatic episodes of vaginosis in women. In a mouse model of Chlamydia infection, we observed preferential expression of CCR2, CCR5, CXCR6, and CD11c. However, during bacterial vaginosis in women, only CCR5 and CD11c were prominently expressed by effector T cells. We also investigated other homing molecules, like α4β1 and α4β7, previously suggested as essential for homing to the genital mucosa. Interestingly, these molecules showed differential expression in the patients. Surprisingly, CD11c, an integrin chain not commonly analyzed in the context of T cell immunity, was consistently elevated in all activated effector CD8+ T cell subsets studied. Additionally, CD11c expression was induced after systemic infection in mice, indicating that it is not exclusive to genital tract infection. Microarray analyses of activated effector T cells expressing CD11c from naïve mice revealed enrichment for genes associated with natural killer cells. Notably, flow cytometry analysis of murine CD11c+ T cells showed markers typically linked with non-conventional T cell subsets, such as γδ T cells and invariant natural killer T cells. However, in women, the CD11c fraction of blood and cervical tissue mainly consisted of γδ T cells and CD8+ T cells. These CD11c+ cells exhibited high activation levels and greater interferon (IFN)-γ secretion compared to CD11c- T cells. Moreover, circulating CD11c+ T cells in women correlated with the expression of multiple adhesion molecules, indicating a high potential for tissue homing. Our findings suggest that CD11c expression distinguishes a specific population of circulating T cells during bacterial infection, displaying innate capacity and mucosal homing potential. Furthermore, its increase in response to genital tract disorders may serve as a novel surrogate marker of mucosal immunity in women, offering potential applications in diagnostics and therapeutics. Further investigation is warranted to explore this potential thoroughly.[cat] Les infeccions de transmissió sexual habituals com la gonorrea, la clamídia, la sífilis i la tricomoniasi són la principal causa de morbiditat als països en desenvolupament. Els adolescents i les dones joves tenen les taxes més altes d’aquestes infeccions. Les infeccions preexistents del tracte genital femení afecten el desenvolupament i la patogènesi d'altres infeccions de transmissió sexual, donant lloc a conseqüències a llarg termini com ara la malaltia inflamatòria pèlvica, el càncer i la infertilitat. Els esforços per desenvolupar vacunes contra les infeccions de transmissió sexual s'han vist obstaculitzats per la incapacitat de mesurar les respostes immunitàries al tracte genital. Les vacunes efectives haurien de generar respostes immunitàries localitzades als llocs d'exposició potencials per proporcionar un control de la infecció més ràpid i robust. Els assajos que es basen en mostres de sang proporcionen una imatge completa del trànsit cel·lular entre els ganglis limfàtics i la mucosa, especialment poc després de la infecció. Estudiar i caracteritzar el perfil de “homing” de les cèl·lules T esdevé crucial per entendre les respostes immunitàries de la mucosa contra les infeccions, a més a més, es necessiten nous mètodes per mesurar i manipular les respostes immunes per desenvolupar vacunes eficaces contra les ITS. En aquest estudi, es va examinar com s'expressen les molècules d'adhesió en les cèl·lules T efectores circulants després de la infecció de la mucosa del tracte genital femení en ratolins i durant episodis simptomàtics de vaginosi en dones. En un model de ratolí d'infecció per Chlamydia, vam observar l'expressió preferent de CCR2, CCR5, CXCR6 i CD11c. Tanmateix, durant la vaginosi bacteriana en les dones, només CCR5 i CD11c es van expressar de manera destacada per les cèl·lules T efectores. També vam investigar altres molècules d'homing, com α4β1 i α4β7, suggerides anteriorment com a essencials per a l'homing a la mucosa genital. Curiosament, aquestes molècules van mostrar una expressió diferencial en els pacients. Sorprenentment, CD11c, una cadena d'integrines que no s'analitza habitualment en el context de la immunitat de les cèl·lules T, es va elevar constantment en tots els subconjunts de cèl·lules T CD8+ efectores i activades. A més, es va induir l'expressió CD11c després d'una infecció sistèmica en ratolins, cosa que indica que no és exclusiva de la infecció del tracte genital. Les anàlisis de microarrays de cèl·lules T efectores activades que expressen CD11c de ratolins naïfs van revelar un enriquiment per a gens associats a cèl·lules assassines naturals (NK). En particular, el anàlisi de citometria de flux de cèl·lules T CD11c+ murines va mostrar marcadors típicament vinculats amb subconjunts de cèl·lules T no convencionals, com ara cèl·lules T γδ i cèl·lules T assassines naturals invariants (iNKT). Tanmateix, a les dones, la fracció CD11c de la sang i el teixit cervical consistia principalment en cèl·lules T γδ i cèl·lules T CD8+. Aquestes cèl·lules CD11c+ presentaven alts nivells d'activació i una secreció més gran d'interferó (IFN)-γ en comparació amb les cèl·lules CD11c-T. A més, les cèl·lules T CD11c+ circulants a les dones es van correlacionar amb l'expressió de múltiples molècules d'adhesió, cosa que indica un alt potencial per el “homing” cap a teixit. Els nostres resultats suggereixen que l'expressió CD11c distingeix una població específica de cèl·lules T circulants durant la infecció bacteriana, mostrant una capacitat innata i un potencial de “homing” cap a la mucosa

Adhesion molecules and effector T cells subsets associated with female genital tract infection

[eng] Common sexually transmitted infections like gonorrhea, Chlamydia, syphilis, and trichomoniasis are the leading cause of morbidity in developing countries. Adolescents and young women have the highest rates of STIs in developing countries. Pre-existing female genital tract infections affect the development and pathogenesis of other sexually transmitted infections, leading to long-term consequences such as pelvic inflammatory disease, cancer, and infertility. Efforts to develop vaccines against sexually transmitted infections have been hindered by the inability to measure immune responses in the genital tract. Effective vaccines should generate localized immune responses at potential exposure sites to provide faster and more robust infection control. Assays rely on blood samples to provide a complete picture of cellular traffic between lymph nodes and the mucosa, especially soon after infection. Studying and characterizing the homing profile of T cells becomes crucial to understanding mucosal immune responses against infections, and new methods to measure and manipulate immune responses are needed to develop effective STI vaccines. In this study, we examined how adhesion molecules are expressed by circulating effector T cells following mucosal infection of the female genital tract in mice and during symptomatic episodes of vaginosis in women. In a mouse model of Chlamydia infection, we observed preferential expression of CCR2, CCR5, CXCR6, and CD11c. However, during bacterial vaginosis in women, only CCR5 and CD11c were prominently expressed by effector T cells. We also investigated other homing molecules, like α4β1 and α4β7, previously suggested as essential for homing to the genital mucosa. Interestingly, these molecules showed differential expression in the patients. Surprisingly, CD11c, an integrin chain not commonly analyzed in the context of T cell immunity, was consistently elevated in all activated effector CD8+ T cell subsets studied. Additionally, CD11c expression was induced after systemic infection in mice, indicating that it is not exclusive to genital tract infection. Microarray analyses of activated effector T cells expressing CD11c from naïve mice revealed enrichment for genes associated with natural killer cells. Notably, flow cytometry analysis of murine CD11c+ T cells showed markers typically linked with non-conventional T cell subsets, such as γδ T cells and invariant natural killer T cells. However, in women, the CD11c fraction of blood and cervical tissue mainly consisted of γδ T cells and CD8+ T cells. These CD11c+ cells exhibited high activation levels and greater interferon (IFN)-γ secretion compared to CD11c- T cells. Moreover, circulating CD11c+ T cells in women correlated with the expression of multiple adhesion molecules, indicating a high potential for tissue homing. Our findings suggest that CD11c expression distinguishes a specific population of circulating T cells during bacterial infection, displaying innate capacity and mucosal homing potential. Furthermore, its increase in response to genital tract disorders may serve as a novel surrogate marker of mucosal immunity in women, offering potential applications in diagnostics and therapeutics. Further investigation is warranted to explore this potential thoroughly.[cat] Les infeccions de transmissió sexual habituals com la gonorrea, la clamídia, la sífilis i la tricomoniasi són la principal causa de morbiditat als països en desenvolupament. Els adolescents i les dones joves tenen les taxes més altes d’aquestes infeccions. Les infeccions preexistents del tracte genital femení afecten el desenvolupament i la patogènesi d'altres infeccions de transmissió sexual, donant lloc a conseqüències a llarg termini com ara la malaltia inflamatòria pèlvica, el càncer i la infertilitat. Els esforços per desenvolupar vacunes contra les infeccions de transmissió sexual s'han vist obstaculitzats per la incapacitat de mesurar les respostes immunitàries al tracte genital. Les vacunes efectives haurien de generar respostes immunitàries localitzades als llocs d'exposició potencials per proporcionar un control de la infecció més ràpid i robust. Els assajos que es basen en mostres de sang proporcionen una imatge completa del trànsit cel·lular entre els ganglis limfàtics i la mucosa, especialment poc després de la infecció. Estudiar i caracteritzar el perfil de “homing” de les cèl·lules T esdevé crucial per entendre les respostes immunitàries de la mucosa contra les infeccions, a més a més, es necessiten nous mètodes per mesurar i manipular les respostes immunes per desenvolupar vacunes eficaces contra les ITS. En aquest estudi, es va examinar com s'expressen les molècules d'adhesió en les cèl·lules T efectores circulants després de la infecció de la mucosa del tracte genital femení en ratolins i durant episodis simptomàtics de vaginosi en dones. En un model de ratolí d'infecció per Chlamydia, vam observar l'expressió preferent de CCR2, CCR5, CXCR6 i CD11c. Tanmateix, durant la vaginosi bacteriana en les dones, només CCR5 i CD11c es van expressar de manera destacada per les cèl·lules T efectores. També vam investigar altres molècules d'homing, com α4β1 i α4β7, suggerides anteriorment com a essencials per a l'homing a la mucosa genital. Curiosament, aquestes molècules van mostrar una expressió diferencial en els pacients. Sorprenentment, CD11c, una cadena d'integrines que no s'analitza habitualment en el context de la immunitat de les cèl·lules T, es va elevar constantment en tots els subconjunts de cèl·lules T CD8+ efectores i activades. A més, es va induir l'expressió CD11c després d'una infecció sistèmica en ratolins, cosa que indica que no és exclusiva de la infecció del tracte genital. Les anàlisis de microarrays de cèl·lules T efectores activades que expressen CD11c de ratolins naïfs van revelar un enriquiment per a gens associats a cèl·lules assassines naturals (NK). En particular, el anàlisi de citometria de flux de cèl·lules T CD11c+ murines va mostrar marcadors típicament vinculats amb subconjunts de cèl·lules T no convencionals, com ara cèl·lules T γδ i cèl·lules T assassines naturals invariants (iNKT). Tanmateix, a les dones, la fracció CD11c de la sang i el teixit cervical consistia principalment en cèl·lules T γδ i cèl·lules T CD8+. Aquestes cèl·lules CD11c+ presentaven alts nivells d'activació i una secreció més gran d'interferó (IFN)-γ en comparació amb les cèl·lules CD11c-T. A més, les cèl·lules T CD11c+ circulants a les dones es van correlacionar amb l'expressió de múltiples molècules d'adhesió, cosa que indica un alt potencial per el “homing” cap a teixit. Els nostres resultats suggereixen que l'expressió CD11c distingeix una població específica de cèl·lules T circulants durant la infecció bacteriana, mostrant una capacitat innata i un potencial de “homing” cap a la mucosa

Expression of CD11c Is Associated with Unconventional Activated T Cell Subsets with High Migratory Potential

Ajudes rebudes: Marie Curie Career Integration Grant; Dexeus Foundation for Women's Health Research; i Contratos Ramón y CajalCD11c is an α integrin classically employed to define myeloid dendritic cells. Although there is little information about CD11c expression on human T cells, mouse models have shown an association of CD11c expression with functionally relevant T cell subsets. In the context of genital tract infection, we have previously observed increased expression of CD11c in circulating T cells from mice and women. Microarray analyses of activated effector T cells expressing CD11c derived from naïve mice demonstrated enrichment for natural killer (NK) associated genes. Here we find that murine CD11c+ T cells analyzed by flow cytometry display markers associated with non-conventional T cell subsets, including γδ T cells and invariant natural killer T (iNKT) cells. However, in women, only γδ T cells and CD8+ T cells were enriched within the CD11c fraction of blood and cervical tissue. These CD11c+ cells were highly activated and had greater interferon (IFN)-γ secretory capacity than CD11c- T cells. Furthermore, circulating CD11c+ T cells were associated with the expression of multiple adhesion molecules in women, suggesting that these cells have high tissue homing potential. These data suggest that CD11c expression distinguishes a population of circulating T cells during bacterial infection with innate capacity and mucosal homing potential

Pro-Oxidant Activity of Amine-Pyridine-Based Iron Complexes Efficiently Kills Cancer and Cancer Stem-Like Cells.

Differential redox homeostasis in normal and malignant cells suggests that pro-oxidant-induced upregulation of cellular reactive oxygen species (ROS) should selectively target cancer cells without compromising the viability of untransformed cells. Consequently, a pro-oxidant deviation well-tolerated by nonmalignant cells might rapidly reach a cell-death threshold in malignant cells already at a high setpoint of constitutive oxidative stress. To test this hypothesis, we took advantage of a selected number of amine-pyridine-based Fe(II) complexes that operate as efficient and robust oxidation catalysts of organic substrates upon reaction with peroxides. Five of these Fe(II)-complexes and the corresponding aminopyridine ligands were selected to evaluate their anticancer properties. We found that the iron complexes failed to display any relevant activity, while the corresponding ligands exhibited significant antiproliferative activity. Among the ligands, none of which were hemolytic, compounds 1, 2 and 5 were cytotoxic in the low micromolar range against a panel of molecularly diverse human cancer cell lines. Importantly, the cytotoxic activity profile of some compounds remained unaltered in epithelial-to-mesenchymal (EMT)-induced stable populations of cancer stem-like cells, which acquired resistance to the well-known ROS inducer doxorubicin. Compounds 1, 2 and 5 inhibited the clonogenicity of cancer cells and induced apoptotic cell death accompanied by caspase 3/7 activation. Flow cytometry analyses indicated that ligands were strong inducers of oxidative stress, leading to a 7-fold increase in intracellular ROS levels. ROS induction was associated with their ability to bind intracellular iron and generate active coordination complexes inside of cells. In contrast, extracellular complexation of iron inhibited the activity of the ligands. Iron complexes showed a high proficiency to cleave DNA through oxidative-dependent mechanisms, suggesting a likely mechanism of cytotoxicity. In summary, we report that, upon chelation of intracellular iron, the pro-oxidant activity of amine-pyrimidine-based iron complexes efficiently kills cancer and cancer stem-like cells, thus providing functional evidence for an efficient family of redox-directed anti-cancer metallodrugs

Comparison of specific phenotype frequencies based on CD11c expression in blood samples from healthy women.

<p>Comparison of the frequency of CD8, γδTCR, MAIT and iNKT (Vα24) in CD11c⁺ and CD11c^-, CCR7^- CD3⁺ T cells from the same individual. (<b>a</b>) Fresh blood (n = 10) and (<b>b</b>) PBMC (n = 6). Gating strategy consisted on the following consecutive gates: lymphocytes, singlets, live CD14^- CD19^- CD3⁺ T cells and CCR7^- CD11c⁺or CD11c^- T cells (<i>see</i> <b><a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0154253#pone.0154253.s003" target="_blank">S3 Fig</a></b> <i>for further details</i>). Data were analyzed using the paired Student’s t-test.</p

Adhesion Molecules Associated with Female Genital Tract Infection

Altres ajuts: Marie Curie Career Integration Grant i una beca Fundació Dexeus Salut de la DonaEfforts to develop vaccines that can elicit mucosal immune responses in the female genital tract against sexually transmitted infections have been hampered by an inability to measure immune responses in these tissues. The differential expression of adhesion molecules is known to confer site-dependent homing of circulating effector T cells to mucosal tissues. Specific homing molecules have been defined that can be measured in blood as surrogate markers of local immunity (e.g. α4β7 for gut). Here we analyzed the expression pattern of adhesion molecules by circulating effector T cells following mucosal infection of the female genital tract in mice and during a symptomatic episode of vaginosis in women. While CCR2, CCR5, CXCR6 and CD11c were preferentially expressed in a mouse model of Chlamydia infection, only CCR5 and CD11c were clearly expressed by effector T cells during bacterial vaginosis in women. Other homing molecules previously suggested as required for homing to the genital mucosa such as α4β1 and α4β7 were also differentially expressed in these patients. However, CD11c expression, an integrin chain rarely analyzed in the context of T cell immunity, was the most consistently elevated in all activated effector CD8+ T cell subsets analyzed. This molecule was also induced after systemic infection in mice, suggesting that CD11c is not exclusive of genital tract infection. Still, its increase in response to genital tract disorders may represent a novel surrogate marker of mucosal immunity in women, and warrants further exploration for diagnostic and therapeutic purposes