References

Park HB, Kim KH, Kim JH, Kim SI, Oh YM, Kang M, Lee S, Hwang S, Lee H, Lee T, Park S, Lee JE, Jeong GR, Lee DH, Youn H, Choi EY, Son WC, Chung SJ, Chung J, Choi K (2024) Improved safety of chimeric antigen receptor T cells indirectly targeting antigens via switchable adapters. Nat Commun 15(1):9917. doi: 10.1038/s41467-024-53996-7 PMID: 39557825

Objective: To show that switchable CAR-T cells with a tumor targeting adaptor can mitigate on-target off-tumor toxicity against a low selectivity tumor antigen that cannot be targeted by conventional CAR-T cells, such as CD40.

Summary: The system is composed of anti-cotinine murine CAR-T cells and cotinine-labeled anti-CD40 single chain variable fragments (scFv), with which the authors show selective tumor killing while sparing CD40-expressing normal cells including macrophages in a mouse model of lymphoma. The authors evaluated whether Cot CAR-T cells could be depleted by Cot-saporin in vivo in an allogeneic CAR-T cell transfer model. When Balb/C mice transplanted with B6 bone marrow cells were injected with B6 Cot CAR-T cells, the transferred Cot CAR-T cells expanded in the peripheral blood in response to Balb/C alloantigen. However, when Cot-saporin was administered during this expansion phase, the Cot CAR-T cells failed to expand and were subsequently eliminated in the blood. Thus, Cot-saporin-mediated CotCAR-T cell suicide was confirmed in vitro and in vivo.

Usage: in vitro Cot CAR-T cell depletion by cotinine-drug conjugates: Peptides were incubated with saporin-labeled streptavidin (IT-27) at a molar ratio of 4:1 to generate cotinine-saporin conjugate (Cot-saporin). For Cot-saporin-dependent cytotoxicity assays on Cot CAR-T cells, a 1:1 mixed population (50,000 cells each) of Cot CAR-T cells (target cells) and control T cells (bystander non-CAR-T cells) were incubated with various doses of Cot-saporin for 48 h in medium containing human IL-2. Seven days after CAR-T cell transfer, Cot-saporin was administered intraperitoneally three times at 3-day intervals.

Related Products: Streptavidin-ZAP (Cat. #IT-27)

Rehal SK (2024) Investigating the role of the ventral anterior cingulate cortex (vacc) in placebo analgesia. Univ Toronto Thesis.

Objective: To examine whether opioid receptors in the ventral anterior cingulate cortex (vACC) were critical for the placebo response evoked by conditioning with 10 mg/kg morphine.

Summary: Significant differences were observed before and after injection of morphine for Dermorphin-SAP-treated mice throughout the conditioning days. At the same time, a percentage of anti-allodynia remained constant over the conditioning phase. Thus, mice displayed the analgesic effects of morphine during the conditioning phase. Additionally, microinjection of Dermorphin-SAP into the vACC did not influence pharmacological conditioning with morphine as there was no difference between either group during the conditioning phase. Selective ablation of mu opioid-expressing neurons in the vACC via Dermorphin-SAP led to a lack of placebo analgesia.

Usage: Bilateral injections were carried out at a rate of 50 nl/min of Dermorphin-SAP (IT-12) or 200 nl total injection volume of Blank-SAP control (IT-21).

Related Products: Dermorphin-SAP / MOR-SAP (Cat. #IT-12), Blank-SAP (Cat. #IT-21)

Ammar AO (2024) Involvement of peptidergic Edinger-Westphal nucleus in the neurobiology of migraine and acute alcohol exposure. Univ Pecs Thesis.

Objective: To confirm the role of Edinger-Westphal nucleus/urocortin1 (EWcp/UCN1) neurons in migraine. They hypothesized that selective ablation of EWcp/UCN1 neurons will influence the migraine-related behaviors induced by CGRP.

Summary: Upon selective ablation of EWcp/UCN1 neurons, authors examined the migraine-related behaviors in response to calcitonin gene-related peptide (CGRP) treatment. Leptin-SAP treatment significantly reduced the number of UCN1 immunoreactive neurons in the EWcp compared to naïve mice. Before ablation of EWcp/UCN1 neurons, CGRP treatment significantly reduced the periorbital withdrawal threshold compared to saline.

Usage: For selective UCN1 neuron ablation, 50 nl of Leptin-SAP was microinjected into the rostral and caudal parts of the EWcp area.

Related Products: Leptin-SAP (Cat. #IT-47)

Schaible HG, König C, Ebersberger A (2024) Spinal pain processing in arthritis: Neuron and glia (inter)actions. J Neurochem 168(11):3644-3662. doi: 10.1111/jnc.15742 PMID: 36520021

Objective: To address the mechanisms of spinal sensitization evoked by arthritis.

Summary: Neutralization of spinal cytokines by intrathecal interventions attenuates mechanical hyperalgesia. This effect may in part result from local suppression of spinal sensitization and in part from efferent effects which attenuate the inflammatory process in the joint. In summary, arthritis evokes significant spinal hyperexcitability which is likely to contribute to the phenotype of arthritis pain in patients

Usage: Selective microglia destruction with the immunotoxin saporin conjugated to Mac1 antibody (Mac-1-SAP Cat #IT-06, recognizes Mac1 receptor on microglia) attenuated the development of hyperalgesia.

Related Products: Mac-1-SAP mouse/human (Cat. #IT-06)

Kuroda M, Komatsu N, Kosai A, Hamakubo T, Abe T (2024) Anti-EGFR antibody immunotoxins improve cytotoxic effects in the salivary gland cancer A253 cell line. doi: 10.1016/j.ajoms.2024.11.003

Objective: To confirm the antitumor effects of IT-Cetuximab (IT-Cmab= saporin-conjugated anti-EGFR antibody), which is cetuximab conjugated with a toxin, targeting salivary gland cancers—a type of cancer with limited effective treatment options aside from surgery.

Summary: Cmab alone exhibited no cytotoxic effects, but IT-Cmab demonstrated concentration-dependent cytotoxic effects in A253 cells.

Usage: Cytotoxicity assay (@1.34pM to 4.2nM)

Related Products: Saporin (Cat. #PR-01)

Lin CCJ, Tian Y, Tanzi RE, Jorfi M (2024) Approaches for studying neuroimmune interactions in Alzheimer’s disease. Trends Immunol S1471-4906(24)00248-5. doi: 10.1016/j.it.2024.10.002 PMID: 39537528

Objective: To examine cutting-edge strategies – encompassing animal and cellular models – used to investigate the roles of peripheral immune cells in AD.

Summary: Recent studies using rodent models and innovative human-based cellular systems are beginning to shed light on how peripheral immune cells infiltrate the brain and modulate disease progression.

Usage: Strategies for bone marrow depletion using anti-CD45 or anti-CD117 antibodies conjugated with the ribosome-inactivating protein saporin have been used.

Related Products: Anti-CD117-SAP (Cat. #IT-83), Anti-CD45.2-SAP (Cat. #IT-91)

See Also:

• Palchaudhuri R et al. Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin. Nat Biotechnol 34:738-745, 2016.
• Czechowicz A et al. Selective hematopoietic stem cell ablation using CD117-antibody-drug-conjugates enables safe and effective transplantation with immunity preservation. Nat Commun 10:617, 2019.

Malatiali S, Oriowo M (2024) Losartan is more effective than angiotensin (1-7) in preventing thyroxine-induced renal injury in the rat. Thyroid Res 17(1):22. doi: 10.1186/s13044-024-00211-w PMID: 39491028

Objective: To elaborate the role of AngII in thyroxine-induced renal injury and the possible protective role of Ang 1–7.

Summary: Losartan is more protective than captopril against thyroxine-induced renal changes while Ang 1–7 offered no protection.

Usage: Western Blot: 1:500 dilution

Related Products: Angiotensin II receptor (AT-1R) Rabbit Polyclonal, affinity-purified (Cat. #AB-N27AP)

Moreno-Rodríguez M, Martínez-Gardeazabal J, Bengoetxea de Tena I, Llorente-Ovejero A, Lombardero L, González de San Román E, Giménez-Llort L, Manuel I, Rodríguez-Puertas R (2024) Cognitive improvement via cortical cannabinoid receptors and choline-containing lipids. Br J Pharmacol doi: 10.1111/bph.17381 PMID: 39489624

Objective: Authors hypothesized that activation of the endocannabinoid system may confer neuroprotection against cholinergic degeneration.

Summary: Degeneration, induced by 192-IgG-saporin, of baso-cortical cholinergic pathways resulted in memory deficits and decreased cortical levels of lysophosphatidylcholines (LPC). The cannabinoid agonist WIN55,212-2 restored cortical cholinergic transmission and LPC levels via activation of cannabinoid receptors. This activation altered cortical lipid homeostasis mainly by reducing sphingomyelins in lesioned animals. These modifications were crucial for memory recovery.

Usage: Basal forebrain cholinergic degeneration was induced following bilateral stereotaxic injection of 192IgG-saporin (130 ng/μl, IT-01) into the nucleus basalis magnocellularis (NBM).

Related Products: 192-IgG-SAP (Cat. #IT-01)

Chuang KH, Zhou XA, Xia Y, Li z, Qian L, Eeles E, Ngiam G, Fripp J, Coulson EJ (2024) Cholinergic basal forebrain neurons regulate vascular dynamics and cerebrospinal fluid flux. bioRxiv 2024.08.25.609536. doi: 10.1101/2024.08.25.609536

Objective: To show that vascular-CSF coupling correlates with cortical cholinergic activity in non-demented aged humans.

Summary: Waste from the brain is cleared via a cerebrospinal fluid (CSF) exchange pathway. Problems in this pathway is suggested to underlie the pathogenesis of many brain conditions. Cerebrovascula oscillation that couples with pulsatile CSF inflow is suggested to drive the flow of fluid, however how this coupling is regulated in unlcear. The resultsfor the study suggest a neurovascular mechanism by which CSF/glymphatic flux is modulated by cholinergic neuronal activity, thereby providing a conceptual basis for the development of diagnostics and treatments for glymphatic dysfunction.

Usage: Injections of mu-p75-SAP (0.5 mg/ml, IT-16) or control Rabbit-IgG-SAP (0.5 mg/ml, IT-35) were performed into the border between the medial septum and ventral diagonal band. In the first study, the toxin was infused at a rate of 0.4 μl/min (1.5μl total volume), which resulted in a large amount of ablation. In the second study, the toxin concentration was reduced to 0.3 mg/ml to preserve more cholinergic neurons and was infused at a rate of 0.18μl/min (1.0μl total volume).

Related Products: mu p75-SAP (Cat. #IT-16), Rabbit IgG-SAP (Cat. #IT-35)

Blanco T, Nakagawa H, Musayeva A, Krauthammer M, Singh RB, Narimatsu A, Ge H, Shoushtari SI, Dana R (2024) Acquired immunostimulatory phenotype of migratory CD103+ DCs promotes alloimmunity following corneal transplantation. JCI Insight 9(20):e182469. doi: 10.1172/jci.insight.182469 PMID: 39235864

Objective: To investigate the interaction between antigen-presenting cell subsets, specifically CD11b+ dendritic cells (DC2) and CD103+ dendritic cells (DC1),in the context of transplant immunity.

Summary: The findings highlight the critical role of CD103+ DC1 in modulating host alloimmune responses. In recipients with uninflamed corneal beds, migratory CD103+ DC1 exhibit a tolerogenic phenotype. These cells influence the immunogenic behavior of CD11b+ DC2 primarily through IL-10 production, suppressing alloreactive CD4+ Th1 cells via the PD-L1/PD-1 pathway and promoting Treg-mediated tolerance through αvβ8 integrin–activated TGF-β1. Together, these mechanisms contribute to improved graft survival.

Usage: In vivo depletion of CD103+ DC1: Recipient BALB/c or RAG-/- mice were administered 2.0 mg/kg of Anti-CD103-SAP (IT-50) intraperitoneally, or an equivalent dose of control conjugate (IgG-SAP).

Related Products: Anti-CD103-SAP (Cat. #IT-50), Rat IgG-SAP (Cat. #IT-17)