Local Immune Effects Research Articles

Antibody targeting of anaerobic bacteria warms cold tumors and improves the abscopal effect of radiotherapy

BackgroundThe combination of immune checkpoint inhibitors with radiotherapy can enhance the immunomodulation by RT and reduce the growth of distant unirradiated tumors (abscopal effect); however, the results are still not very satisfactory. Therefore, new treatment options are needed to enhance this effect. Our previous study showed that the combination of Bifidobacterium (Bi) and its specific monoclonal antibody (mAb) could target and alleviate hypoxia at the tumor site and act as a radiosensitizer. In this study, we explored the anti-tumor efficacy of quadruple therapy (Bi + mAb and RT + αPD-1). The current study also aimed to probe into the complex immune mechanisms underlying this phenomenon.MethodsConstructed 4T1 breast and CT26 colon cancer tumor models. A comprehensive picture of the impact of constructed quadruple therapy was provided by tumor volume measurements, survival analysis, PET/CT imaging, immune cell infiltration analysis and cytokine expression levels.ResultsThe abscopal effect was further amplified in the “cold” tumor model and prolonged survival in tumor-bearing mice. Bi can colonized in primary and secondary tumors and direct the mAb to reach the tumor site, activate complement, enhance the ADCC effect and initiate the innate immune response. Then combined with αPD-1 and radiotherapy to stimulate adaptive immune response and synergize with cytokines to expand the immune efficacy and generate effective anti-tumor immune response.ConclusionsBi was used as an artificially implanted anaerobic target to cause a transient “infection” at the tumor, causing the tumor to become locally inflamed and “hot”, and at the same time, mAb was used to target Bi to enhance the local immune effect of the tumor, and then combined with radiotherapy and αPD-1 to amplify the abscopal effect in multiple dimensions. Therefore, the present study provided a new idea for the multipotent immune-activating function of antibody-targeted anaerobic bacteria for the RT treatment of extensively metastasized cancer patients.

Changes in cervical cytokine profiles following focused ultrasound treatment for high-risk human papillomavirus infection-related low-grade cervical lesions.

To evaluate the changes of cervical soluble immune markers after focused ultrasound (FU) treatment to explore the underlying local immune effects of FU in the treatment of high-risk human papillomavirus (HR-HPV) infection-related low-grade squamous intraepithelial lesion (LSIL). A total of 35 patients diagnosed with HR-HPV infection-related histological LSIL who met the inclusion criteria were enrolled in this prospective study and treated with FU. The authors used cytometric bead array to measure T-helper type 1 (Th1) cytokine (interleukin [IL] 2, tumor necrosis factor, and interferon γ) and Th2 cytokine (IL-4, IL-5, IL-6, and IL-10) levels in the cervicovaginal lavage of patients before and 3 months after FU treatment. After FU treatment, the concentrations of Th2 cytokines IL-5 and IL-6 were significantly lower than those before FU treatment (P = 0.044 and P = 0.028, respectively). HR-HPV infection was cleared in 27 patients, with a clearance rate of 77.1% (27 of 35). The concentration of IL-4 in patients with HR-HPV clearance after FU treatment was significantly lower than that in patients without HR-HPV clearance (P = 0.045). FU can inhibit the production of certain Th2 cytokines and may improve the local immune status of the cervix, thereby eliminating HR-HPV infection.

Metformin generates profound alterations in systemic and tumor immunity with associated antitumor effects

BackgroundMetformin is a commonly used antidiabetic medication which has demonstrated promise as an anticancer agent alone and in combination with conventional treatment regimens. There is increasing evidence that metformin can...

Objective measurement and clinical significance of TILs in non-small cell lung cancer.

Tumor-infiltrating lymphocytes (TILs) are usually measured using subjective methods. Studies suggest that TIL subtypes have independent roles in cancer and that they could support the use of novel immunostimulatory therapies. We simultaneously measured TIL subtypes in non-small cell lung cancer (NSCLC) samples using objective methods and determined their relationship with clinico-pathologic characteristics and survival. Using multiplexed quantitative fluorescence (QIF), we measured the levels of CD3, CD8, and CD20 in 552 NSCLC from two independent collections represented in tissue microarrays (YTMA79, n = 202 and YTMA140, n = 350). The level of TILs was obtained in different tumor compartments using cytokeratin stain to define tumor cells and 4',6-Diamidino-2-Phenylindole. Association of TILs with clinical parameters was determined using univariate and multivariable analyses. All statistical tests were two-sided. In both NSCLC collections there was a low correlation between the three TIL markers (linear regression coefficients (R(2)) = 0.19-0.22, P < .001 for YTMA79 and R(2) = 0.23-0.32, P < .001 for YTMA140). No consistent association between the level of TIL subtypes and age, sex, smoking history, tumor size, stage, and histology type was found. In univariate analysis, an elevated CD3 or CD8 signal was statistically significantly associated with longer survival in both collections. However, only CD8 was independent from age, tumor size, histology, and stage in multivariable analysis. High CD20 was associated with longer survival in the YTMA79 cohort. Increased levels of CD3 and CD8 + TILs are associated with better outcome in NSCLC, but only CD8 is independent from other prognostic variables. Objective measurement of TIL subpopulations could be useful to predict response or evaluate the local immune effect of anticancer immune checkpoint inhibitors.

Route-dependent systemic and local immune effects following exposure to solutions prepared from titanium dioxide nanoparticles

Nanoparticle titanium dioxide (nano-TiO2) is a white pigment widely used in foods, sunscreens, and other cosmetic products. However, it remains unclear whether exposure to nano-TiO2 results in immunosuppressive effects or induces a contact hypersensitivity response. To address these data gaps, studies were conducted with the hypothesis that nano-TiO2 exposure could alter immune responses. After 28 days of oral gavage, nano-TiO2 (1.25–250 mg/kg in 0.5% methylcellulose) produced no significant effects on innate, humoral, or cell-mediated immune functions in female B6C3F1 mice. Furthermore, there were no effects on the weights of selected organs (spleen, thymus, liver, lung, and kidneys with adrenals). Following dermal exposure on the ears for 3 days, nano-TiO2 (2.5–10% w/v in 4:1 acetone:olive oil) did not affect auricular lymph node cell proliferation, although an irritancy response was observed following treatment with 5% and 10% nano-TiO2. Dermal sensitization (2.5–10%) on the back and subsequent challenge (10%) on the right ear with nano-TiO2 produced no significant effects on percentage ear swelling in the Mouse Ear Swelling Test (MEST). However, when nano-TiO2 was injected subcutaneously along the mid-line on top of the head at 125–250 mg/kg (in 0.5% methylcellulose), significant increases in auricular lymph node cell proliferation resulted. These results demonstrate that immune effects of nano-TiO2 exposure are route-of-exposure dependent, and they suggest that irritancy and/or potential hypersensitivity responses may occur following parenteral exposure or dermal administration of nano-TiO2 to compromised skin.

B7x deficiency exacerbates renal injury in antibody mediated nephritis (60.7)

Abstract B7x (B7-H4, B7S1) is an important member of the B7/CD28 superfamily of T cell costimulatory proteins. B7x is a negative regulator of CD4+ and CD8+ T cell activation and proliferation, and is expressed in both hematopoietic and non-hematopoietic tissues. We hypothesized that B7x is instrumental in the pathogenesis of antibody mediated nephritis. To examine this question we induced nephritis in B7x-deficient (B7x-/-) (n=10) and WT B6 mice (n=10) by passive transfer (i.v.) of nephritogenic antibodies. Following induction of nephritis, we found a marked increase in the levels of serum IgG (p=0.023), IgG2b (p=0.055), and IgG1 (p=0.064) in B7x-/- mice compared to WT mice. Moreover, renal damage was more severe in B7x deficient mice. Serum creatinine and BUN were elevated in the B7x-/- group compared to the WT mice. Furthermore, B7x-/- mice demonstrated worse kidney histopathology, with an increase in infiltrating PMN cells in glomeruli (p&amp;lt;0.0001) and interstitium (p=0.004), crescent formation, and tubular atrophy/dilatation. Immunohistochemical studies revealed increased glomerular infiltration of CD3+ T cells (p=0.034) and CD68+ macrophages (p=0.003) in B7x-/- mice kidneys. At the mRNA level, IL-23 was significantly upregulated in kidneys of B7x-/- mice. Thus, B7x plays a protective role in the pathogenesis of antibody mediated nephritis. The relative contributions of systemic versus local immune effects of B7x in nephrotoxic nephritis are currently being investigated.

Prospective oral food challenge study of two soybean protein isolates in patients with possible milk or soy protein enterocolitis.

The soybean protein isolate used in powdered soybean formula is hydrolyzed more extensively than the isolate which is used in liquid soybean formula in most commercial soybean formulas. Previous in vitro studies have shown differences in human antibody response to these soybean protein isolates. Therefore, a prospective clinical study was undertaken to determine if there were differences in adverse reaction rates to these soybean protein isolates. Forty-three patients with possible milk- and/or soy-protein enterocolitis were enrolled in this study. Patients had 3 separate oral food challenges; using milk formula, soybean powder formula and soybean liquid formula. Ten (23%) patients challenged with milk had positive challenges. Fourteen (33%) patients challenged with powdered soy formula had positive challenges while thirteen (30%) challenged with liquid soy formula had positive challenges. In the 10 patients with positive milk challenges, 6 (60%) had a positive soy challenge. In the group with positive soy challenges, 5 reacted to the powdered soy challenge done first, but not the second challenge with the liquid soy formula, and 4 patients reacted to the liquid soy formula challenge done first, but not the second challenge with the powdered soy formula. These results indicate that a significant number of patients with milk protein enterocolitis have soy protein enterocolitis. In addition, an order effect can be demonstrated in the soy challenges because of the tendency to react to the first soy challenge regardless of the type of isolate. These results suggest that a local immune effect caused by the protein may be present.

Generation of suppressor T cells after local immunization with histocompatibility antigens

Subcutaneous (sc) hind-foot immunization (HFI) of mice with allogeneic spleen cells can induce a state of delayed-type hypersensitivity (DTH) as well as a state of suppression of DTH. This paper deals with the suppression induced by HFI. The state of suppression could be adoptively transferred by spleen cells and lymph node cells between Days 3 and 7 after HFI only. However, in the hind-foot-immunized mice the state of suppression lasted at least 25 days. The suppressor cells expressed the Thy-1 +, Lyt-1 −2 + phenotype and suppressed DTH antigen-specifically. The suppressor cells, however, also suppressed DTH responses to unrelated third-party alloantigens, provided the latter were administered during the induction of DTH together with the same alloantigens that were used for HFI. The HFI-induced T-suppressor cells suppressed the induction phase of DTH (i.e., the proliferative activity of the draining lymph node cells after secondary sc immunization), but not the expression phase of DTH (i.e., the activity of previously activated DTH effector T cells). H-2D compatibility between the donors of the HFI-induced T-suppressor cells and the recipients was required for the adoptive transfer of suppression. The differences in effect of local immunization versus systemic immunization on the induction and functional activity of T-suppressor cells are discussed.

The Effect of Local Immunization in Endolymphatic Sac on the Production of Type II Collagen (cII) Induced Endolymphatic Hydrops

The Effect of Local Immunization in Endolymphatic Sac on the Production of Type II Collagen (cII) Induced Endolymphatic Hydrops

205 The effect of local immunization in endolymphatic SAC (ELS) on the production of type II (CII) collagen induced ear disease (CIAED)

205 The effect of local immunization in endolymphatic SAC (ELS) on the production of type II (CII) collagen induced ear disease (CIAED)

Effects of local immunization of hamsters with glucosyltransferase antigens on infection with Streptococcus sanguis.

The effects of immunization with antigens of the Streptococcus mutans glucosyltransferase (GTF) complex on oral challenge with two Streptococcus sanguis strains (H7PR3 and 34) in hamsters were studied. Antisera to S. mutans GTF complex were able to inhibit one-third (strain H7PR3) to one-half (strain 34) of the S. sanguis GTF activity which could be inhibited when these S. sanguis GTFs were incubated with antisera to S. sanguis GTF. Washed, intact cells of strains H7PR3 and 34 were able to remove a significant amount of enzyme inhibitory activity and immunoglobulin G antibody activity from antisera to S. mutans GTF. These results established the existence of an antigenic relationship between S. sanguis and S. mutans GTFs. The effect of injection of S. mutans strain 6715 GTF or phosphate-buffered saline, incorporated into complete Freund adjuvant, on oral challenge with S. sanguis was compared in 243 hamsters in nine experiments. Salivary and serum GTF inhibitory activity was present in all GTF-injected animals before challenge. After a 2- or 3-day challenge with S. sanguis H7PR3 (seven experiments) or 34 (two experiments), fewer bacteria were recovered from GTF-injected hamsters in every experiment. Significant differences were observed in six of the nine experiments. During the 7- to 21-day period after challenge, 33% of the phosphate-buffered saline-injected sham group (group I) still had S. sanguis recoverable from the molar surfaces, whereas only 19% of the S. mutans GTF-injected group (group II) remained infected with S. sanguis (P less than 0.01). These results suggest that immunization with GTF from S. mutans may influence the colonization potential of S. sanguis in the oral cavity.

Effects of local immunization with glucosyltransferase on colonization of hamsters by Streptococcus mutans.

Experiments were performed to study the effect of antibody to Streptococcus mutans glucosyltransferase (GTF) on the implantation of these organisms in hamsters. Salivary (immunoglobulin A) and serum (immunoglobulin G) antibodies to GTF and GTF-inhibiting activity were elicited by injection of GTF in Freund complete adjuvant in the salivary gland region. Sham-immunized and GTF-immunized groups were then orally challenged with approximately 10(7), 10(8), or 10(9) colony-forming units of cariogenic S. mutans 6715. The results were evaluated by systematically swabbing molars 4 days and approximately 4 weeks after challenge. In general, fewer GTF-immunized hamsters became infected with S. mutans after challenge with 10(7) or 10(8) organisms than did identically challenged sham-immunized hamsters. Of the animals that did become infected, fewer S. mutans colony-forming units were recovered from GTF-immunized hamsters. These results indicate that the presence of antibody to GTF can diminish the ability of S. mutans to implant in the oral cavity of immunized hamsters.

Effects of local immunization of hamsters with glucosyltransferase antigens from Streptococcus sanguis on dental caries caused by Streptococcus mutans

Glucosyltransferase (GTF) from Strep. sanguis strain 34 was tested for antigenic relatedness to GTF from Strep. mutans and for the effect of immunization with this GTF on experimental dental caries caused by Strep. mutans. The sera and salivas of hamsters immunized in the vicinity of the salivary glands with Strep. sanguis strain 34 GTF contained high levels of antibody and GTF-inhibitory activity to the homologous GTF as well as low but significant levels of antibody activity to GTF from Strep. mutans strains E49 (serotype a) and strain 6715 (serotype g). Likewise, sera or salivas from hamsters immunized with either Strep. mutans GTF showed significant reaction with Strep. sanguis GTF in the enzyme-linked immunosorbent assay (ELISA). However, groups of hamsters locally immunized with GTF from Strep. sanguis strain 34, followed by infection with cariogenic Strep. mutans strain E49, showed no protective effect on molar tooth surfaces as measured by extent of infection, caries or lesions compared with sham-immunized groups. Control groups immunized with Strep. mutans strain 6715 GTF and infected with Strep. mutans strain E49 organisms had significantly reduced caries and numbers of lesions ( p < 0.02) compared with sham-immunized groups. Therefore, the ability of GTF antigens to elicit a protective immune response among strains of Strep. mutans did not extend to the use of GTF from Strep. sanguis strain 34 as antigen under the conditions of these experiments.

Effects of local immunization with glucosyltransferase fractions from Streptococcus mutans on dental caries in hamsters caused by homologous and heterologous serotypes of Streptococcus mutans.

Seven serotypes of Streptococcus mutans have been identified. The biochemical, genetic, and serological characteristics of these serotypes have indicated that certain serotypes are quite similar, whereas others are quite distinct. The effect of local immunization with glucosyltransferase (GTF) enzymes from serotypes a, c, or g on infection and disease caused by homologous or heterologous cariogenic S. mutans is reported. Organisms with either similar (a and g) or different (c and g) biochemical and serological characteristics were selected for heterologous challenge. NIH white hamsters were injected four times at weekly intervals with GTF prepared by 6 M guanidine-hydrochloride elution from water-insoluble glucan of serotypes a, c, or g, which resulted in enzyme (homologous) inhibitory activity in sera and salivas. After infection of GTF-immunized and sham-immunized groups of hamsters with cariogenic S. mutans of the same serotype as the injected antigen (homologous infection) or with S. mutans of a different serotype from the injected antigen (heterologous infection), the numbers of streptomycin-labeled S. mutans, caries, and lesions were determined. Immunization with GTF preparations from each of the three serotypes resulted in statistically significant reductions in the extent of infection and disease and number of lesions caused by infections with homologous cariogenic S. mutans. Statistically significant reductions in these three parameters were also observed in groups immunized with enzyme from serotype a (strain E49) and challenged with cariogenic serotype g (strain 6715) organisms; or immunized with enzyme from serotype c (strain Ingbritt) and challenged with cariogenic serotype g (strain 6715) organisms; or immunized with enzyme from serotype g (strain 6715) and challenged with cariogenic serotype c (strain Ingbritt) organisms. These studies suggest that soluble antigen preparations containing GTF from one serotype may elicit a protective immune response against infection with cariogenic S. mutans from many or possibly all serotypes.

Effects of Local Immunization with Glucosyltransferase Fractions from Streptococcus Mutans on Dental Caries in Rats and Hamsters

The effect of local immunization with glucosyltransferase enzymes of Streptococcus mutans on dental caries in conventional rats, hamsters, and gnotobiotic rats was studied. Injection of these animals with crude or defined glucosyltransferase enzyme preparations incorporated into complete Freund's adjuvant consistently resulted in the presence of antibody in saliva demonstrable by functional inhibition of enzymatic activity and binding of radioactive enzyme. Serum antibody was also present. Two experiments have been completed in conventional rats and one in gnotobiotic rats in which the animals were immunized with crude enzyme antigens. The immunized groups of animals always had lower mean caries scores than comparably sham-immunized or nonimmunized control groups. Hamsters immunized with a defined enzyme preparation, containing no more than three antigenic components (two of which were enzyme), also demonstrated significant reductions in mean caries scores. The numbers of lesions were also always lower in immunized animals. In some cases there were reductions in the numbers of S. mutans that could be recovered from the teeth of immunized, infected animals. The reductions in dental caries and lesions were greater on smooth dental surfaces than on occlusal surfaces, which might be explained as interference with adherence phenomena demonstrated by S. mutans. It is proposed that antibody interference affects dental caries caused by this organism.

Effects of Local Immunization with Streptococcus mutans on Induction of Salivary Immunoglobulin A Antibody and Experimental Dental Caries in Rats

The effect of local immunization with Streptococcus mutans on dental caries in conventional and gnotobiotic rats was studied. Injection of these animals with S. mutans strain 6715 incorporated into complete Freund adjuvant consistently resulted in the presence of antibody in saliva directed to this organism. This antibody was primarily of the immunoglobulin A class as demonstrated by specific antiglobulin augmentation and gel filtration of antibody activity. Serum antibody was also present. Five experiments have been completed in conventional rats and two in gnotobiotic animals. The immunized group of animals always had lower mean caries scores than comparably sham-immunized or nonimmunized control groups. The numbers of lesions were also always lower in the immunized animals, suggesting a possible interference with the formation of new lesions in immunized animals. The reductions in dental caries and lesions were greater on smooth surfaces than on occlusal surfaces. which might be explained as interference with adherence phenomena demonstrated by S. mutans. It is proposed that salivary immunoglobulin A antibody may be viewed as an ecological determinant in the oral cavity by affecting oral microorganisms and possibly their by-products.