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PDF file - 278K, Figure 1. Local low-dose anti-CD40 therapy generates distant anti-tumor effects and inhibits B16 tumor growth.

PDF file - 195K, Figure 3. In vivo biodistribution of target organs 48h and 72h post injection of 125I-CD40-specific and 131I-rat IgG2a control antibody.

Immunotherapy with intratumoral injection of adenoviral vectors expressing CD40L has yielded positive results in experimental and clinical bladder cancer. We therefore hypothesized that anti-CD40 antibody would be effective in this setting. Agonistic CD40 antibodies were developed as vaccine adjuvants but have later been used as treatment of advanced solid tumors and hematologic cancers. Systemic anti-CD40 therapy has been associated with immune-related adverse events, such as cytokine release syndrome and liver toxicity, and local delivery is an attractive approach that could reduce toxicity. Herein, we compared local and systemic anti-CD40 antibody delivery to evaluate efficacy, toxicity, and biodistribution in the experimental MB49 bladder cancer model. Antitumor effects were confirmed in the B16 model. In terms of antitumor efficacy, local anti-CD40 antibody stimulation was superior to systemic therapy at an equivalent dose and CD8 T cells were crucial for tumor growth inhibition. Both administration routes were dependent on host CD40 expression for therapeutic efficacy. In vivo biodistribution studies revealed CD40-specific antibody accumulation in the tumor-draining lymph nodes and the spleen, most likely reflecting organs with frequent target antigen-expressing immune cells. Systemic administration led to higher antibody concentrations in the liver and blood compared with local delivery, and was associated with elevated levels of serum haptoglobin. Despite the lack of a slow-release system, local anti-CD40 therapy was dependent on tumor antigen at the injection site for clearance of distant tumors. To summarize, local low-dose administration of anti-CD40 antibody mediates antitumor effects in murine models with reduced toxicity and may represent an attractive treatment alternative in the clinic. Cancer Immunol Res; 2(1); 80–90. ©2013 AACR.

PDF file - 68K, Table 3: In vivo biodistribution 48h (also see Supplementary Fig. S3A). Statistical differences calculated between anti-CD40 and rat IgG2a and the two administration routes (n=4, Student?s paired t-test).

PDF file - 68K, Table 1: In vivo biodistribution 4h (also see Fig. 4A). Statistical differences calculated between anti-CD40 and rat IgG2a and the two administration routes (n=4, Student?s paired t-test).

PDF file - 67K, Table 2: In vivo biodistribution 24h (also see Fig. 4B). Statistical differences calculated between anti-CD40 and rat IgG2a and the two administration routes (n=4, Student?s paired t-test).

PDF file - 404K, Figure 2. Retained CD40 specificity after radiolabeling and modulation of CD40 expression as well as immune cell distribution after repeated injections of FGK4.5 in vivo.

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Supplementary Methods

Supplementary Table 1. Selection strategy Supplementary Table 2. Kinetic parameters for ADC-1013 and B44 measured by Biacore Supplementary Table 3. In vitro potency (EC50) of ADC-1013 in DC and B cell assays Supplementary table 4. Phenotypic profiling of hCD40tg mice

Purpose: Local administration of immune-activating antibodies may increase the efficacy and reduce the immune-related adverse events associated with systemic immunotherapy of cancer. Here, we report the development and affinity maturation of a fully human agonistic CD40 antibody (IgG1), ADC-1013.Experimental Design: We have used molecular engineering to generate an agonistic antibody with high affinity for CD40. The functional activity of ADC-1013 was investigated in human and murine in vitro models. The in vivo effect was investigated in two separate bladder cancer models, both using human xenograft tumors in immune deficient NSG mice and using a syngeneic bladder cancer model in a novel human CD40 transgenic mouse.Results: Activation of dendritic cells (DC) by ADC-1013 results in upregulation of the costimulatory molecules CD80 and CD86, and secretion of IL12. ADC-1013 also activates DCs from human CD40 transgenic mice, and peptide-pulsed and ADC-1013–stimulated DCs induce antigen-specific T-cell proliferation in vitro. In vivo, treatment with ADC-1013 in a syngeneic bladder cancer model, negative for hCD40, induces significant antitumor effects and long-term tumor-specific immunity. Furthermore, ADC-1013 demonstrates significant antitumor effects in a human bladder cancer transplanted into immunodeficient NSG mice.Conclusions: Our data demonstrate that ADC-1013 induces long-lasting antitumor responses and immunologic memory mediated by CD40 stimulation. To the best of our knowledge, ADC-1013 represents the first immunomodulatory antibody developed for local immunotherapy of cancer. Clin Cancer Res; 21(5); 1115–26. ©2014 AACR.See related commentary by Dronca and Dong, p. 944

Purpose: Immunotherapy with Bacillus Calmette-Guerin (BCG) instillation is recommended for high-risk, non–muscle invasive bladder cancer. Bacillus Calmette-Guerin is not effective in advanced tumors, and better alternatives are warranted. Immunostimulating gene therapy with adenoviral vectors expressing CD40 ligand (AdCD40L) has shown efficacy in tumor models. CD40 ligand stimulates systemic immunity and may be effective in local and invasive human disease.Experimental Design: Patients with invasive bladder cancer scheduled for cystectomy or patients with Ta tumors were enrolled in a phase I/IIa trial. Patients were treated with three cycles of intrabladder Clorpactin WCS-90 prewash, followed by AdCD40L instillation 1 week apart. Safety, gene transfer, immune effects, and antitumor responses were monitored.Results: All eight recruited patients were treated as scheduled, and therapy was well tolerated. The main adverse effect was transient local pain during prewash. Postoperatively, urinary tract infections and one case of late septicemia with elevated potassium were reported. No adverse events were ascribed to vector therapy. Gene transfer was detected in biopsies, and bladders were heavily infiltrated with T cells. The effector marker IFN-γ increased in biopsies, whereas levels of circulating T regulatory cells were reduced. Histologic evaluation indicated that AdCD40L therapy reduced the load of malignant cells.Conclusions: To our knowledge, this is the first report on immunogene therapy in bladder cancer and the first using AdCD40L in vivo. Local AdCD40L gene therapy was safe, boosted immune activation, and should be further evaluated as a single or an adjuvant therapy for urothelial malignancies. Clin Cancer Res; 16(12); 3279–87. ©2010 AACR.

Supplementary Figure 3. Anti-tumor effect on MB49 tumors. ADC-1013 generates a significant anti-tumor effect on CD40 negative MB49 cells in hCD40tg mice. Treatment with ADC-1013 or isotype control was performed on day 7 and 10 post tumor inoculation. Increasing doses of ADC-1013 were administred peritumorally in 100 ul PBS and the individual tumor growth curves are displayed.

Supplementary Figure 2. Microphotographs from cryo sections showing representative images of A) Spleen, B) Liver and C) Brain (one slide representing data from one female and one male mice) from hCD40+ mice stained with an anti-human CD40 antibody or anti mouse CD40 antibody.

Supplementary Figure 1. A) Annexin V positive moDC after ADC-1013 stimulation and B) the effect of cross-linking on percentage of Annexin stained cells (n=6).

Supplementary Data from AdCD40L Immunogene Therapy for Bladder Carcinoma—The First Phase I/IIa Trial

Supplementary Figure 6. Splenocytes from complete responders (CR) from ADC-1013 treatment of MB40 tumors were transplanted to naive mice. The receiving naïve mice were then challenged with MB49 tumors and the tumor volume followed over time until the first mice was sacrificed (**p

Supplementary Figure 5. hCD40tg mice with one tumor on each flank were treated peritumorally in the tumor on the right flank. Tumor volumes of individual untreated tumors (red) and treated tumors (black) are displayed.

Supplementary Figure 4. Anti-tumor effect on MB49 tumors. ADC-1013 generates a significant anti-tumor effect on CD40 negative MB49 cells in hCD40tg mice. Treatment with ADC-1013 or isotype control was performed on day 7 and 10 post tumor inoculation. Increasing doses of ADC-1013 were administred peritumorally in 100 ul PBS and the individual tumor growth curves are displayed.

Tutkimuksessa tutkittiin työssä oppimista kohdeyrityksen yhdessä toimipisteessä. Työelämän muutokset koskettavat yksilöitä, tiimejä ja organisaatiota. On myös esitetty, että formaali koulutusympäristö enää olisi ainut työhön liittyvän oppimisen paikka, vaan että työpaikoista tulee yhä enenevissä määrin oppimisympäristöjä. Tutkimuksen tarkoitus oli tuottaa tutkimuksen kohteena olevalle organisaatiolle työssä oppimiseen liittyvää tietoa, jonka avulla se voi varmistaa ja mahdollistaa yksilöiden ja koko organisaation toimintakyvyn työn muutoksissa. Tutkimme yksilötason ja sosiaalisia oppimisprosesseja, johtamista, oppimisympäristöä ja työpaikan kulttuuria. Tutkimuksen kohteena oli globaalin logistiikkayrityksen 10-henkinen toimihenkilötiimi Suomessa. Tutkimus on kvalitatiivinen. Tutkimuksen aineisto kerättiin yksilö- ja ryhmähaastatteluna. Yksilö- ja ryhmähaastatteluun osallistuivat samat henkilöt, sillä erotuksella, että kahta yksilöhaastattelussa ollutta henkilöä ei otettu mukaan ryhmähaastatteluun valta-aseman vuoksi. Haastattelut toteutettiin COVID-19-viruksen aiheuttaman pandemian vuoksi etähaastatteluna Teams-videoyhteydellä. Haastattelut olivat puolistrukturoituja teemahaastatteluja. Haastatteluaineisto analysoitiin hyödyntäen aineistolähtöistä sisällönanalyysia sekä teemoittelua. Tulosten perusteella voidaan esittää, että tutkimuksen kohteena olleen tiimin työssä oppimisen tärkeimmiksi mahdollistajiksi nousivat oppiminen sosiaalisena tapahtumana, psykologisesti turvallinen työkulttuuri, työntekijöiden kasvamisen ja autonomian salliva johtaminen sekä bottom up - organisaatiomalli. Työssä oppimiseen vaikutti myös se, miten työympäristö oli rakentunut. Tutkimuksella on kolme päämerkitystä. Ensimmäisenä merkityksenä voidaan pitää yksilön sekä tiimin hyvän työssä oppimisen mahdollistavien tekijöiden löytymistä. Toinen merkitys on se, että tutkimuksen perusteella saatiin uusia ehdotuksia koko organisaation käyttöön. Esimerkiksi se, että tiimit voivat tehdä itse työvuorosuunnittelun. Oppiminen näyttäytyi vahvasti sosiaalisena tapahtumana. Kolmantena merkityksenä on uuden pohdinnan tuottaminen. On hyvä miettiä sitä, miten organisaatiot pystyvät toteuttamaan ja turvaamaan sosiaalisen lähioppimisen mahdollisuuksia myös erilaisissa tulevissa haasteellisissa olosuhteissa.