Jordan
 
S.
 
Orange
MD, PhD
Email: 
orange@email.chop.edu
Address: 
Children's Hospital of Philadelphia Abramson Research Center 907A 3516 Civic Center Blvd.
(267) 426-5622
Affiliations
Expertise

Genetic immunodeficiency disorders
Natural Killer cell deficiencies

Dr. Orange is interested in determining the role of natural killer cells in human host defense through the study of inborn genetic errors that affect natural killer cell function. The laboratory has identified a specific functional defect in NK cells of patients with the Wiskott-Aldrich syndrome and has recently opened a phase-I clinical trial based upon these findings (www.wastherapy.com).

Natural killer (NK) cells are lymphocytes critical to host defense that play important roles in surveillance of tumor cells as well as in control of viral infections. They do not undergo genetic recombination to attain specificity and therefore are part of the innate immune system. NK cells mediate cytotoxicity by extruding secretory lysosomes in a directed manner after a favorable balance between the ligation of activating and inhibiting receptors has been achieved. The foundations of NK cell activities and regulation therefore lie at the interface between NK cells and cells with which they are interacting. Molecules accumulate in this region and result in a dynamic structure called the NK cell immunologic synapse (NKIS).

My laboratory is investigating the formation, function and regulation of the NKIS. We have focused upon the cytoskeleton as a critical juncture for these processes due to an interest in a human disease that impairs cytoskeletal function called the Wiskott-Aldrich syndrome. Using cells from patients with this disorder, as well as various cytoskeletal inhibitors, we have shown that the activating NKIS is actin-dependent. This is in contrast to the inhibitory NKIS, which is actin-independent. We have also identified sequential steps required for creation of the activating NKIS and have demonstrated that actin reorganization precedes and is required for microtubular function at the synapse. The microtubules are then needed to translocate lytic granules to the center of the NKIS called the central supramolecular activation cluster (cSMAC). Most recently our work has focused upon cytoskeletal events critical in forming the activating NKIS and we have been evaluating actin complex-associated proteins for their role in granule localization to the cSMAC. We are additionally studying how molecular rearrangement at the NKIS results in activation-induced transcriptional regulation and how this ultimately affects cytotoxic function. NK cells are a most useful model for these studies because they have an easily defined function and a more gradual activation process attributed to the interplay of activating and inhibitory receptors.

Research:

Natural killer (NK) lymphocytes are critical to host defense both for surveillance of tumor cells as well as for control of viral infections. As members of the innate immune system, they do not undergo genetic recombination to attain specificity.  The foundations of NK cell activities and regulation lie at the interface between NK cells and cells with which they interact. Once a favorable balance between the ligation of activating and inhibiting receptors has been achieved, NK cells kill target cells by extruding secretory lysosomes. The dynamic structure at the NK/target cell interface is known as the NK cell immunologic synapse (NKIS).
The focus of work in the Orange laboratory is to better understand the formation, function and regulation of the NKIS.  Using cells from patients with Wiskott-Aldrich syndrome, a human disease featuring impaired cytoskeletal function, we have shown that the activating NKIS is actin-dependent. This is in contrast to the inhibitory NKIS, which is actin-independent. We have also identified sequential steps required for the creation of the activating NKIS and have demonstrated that actin reorganization precedes and is required for microtubule function at the synapse. Our most recent work has focused upon cytoskeletal events required for polarization of the microtubule organizing center and lytic granules to the NKIS, as well as the final translocation of lytic granules to the synaptic membrane.
Lab expertise and resources:

Quantitative confocal microscopy
Colocalization studies
Nuclear localization studies
Dynamic object tracking
Total internal reflection fluorescence microsopy
Imaging under flow stress

IMIG Collaborations:

Orange and Reiner: evaluation and measurement of asymmetry in immune cells



Lab members:


Pinaki Banerjee – Research Associate

Quantitative imaging, fixed cell confocal microscopy
pinaki@email.chop.edu



Sumita Roy Ghanta – Research Associate

Sumita.Roy-Ghanta@uphs.upenn.edu



Jennifer Grier– Graduate student

Quantitative imaging, fixed cell confocal microscopy
grierjt@mail.med.upenn.edu



Emily Mace - Post-doctoral Fellow

Saumya Maru – Undergraduate

saumya.maru@gmail.com



Ashley Mentlik – Graduate student

Quantitative imaging, live cell confocal microscopy
amentlik@mail.med.upenn.edu



Linda Monaco-Shawver – Research Technician

Retroviral transduction
shawver@email.chop.edu



Rahul Pandey – Post-doctoral Fellow

pandeyr@email.chop.edu



Greg Rak – Graduate student

Total internal reflection fluorescence microscopy
grak@vet.upenn.edu



Keri Sanborn – Graduate student

Quantitative imaging, live cell confocal microscopy, flow chamber microscopy experiments
ksanborn@gmail.com

Research Interests
Directed secretion at the cytolytic immunological synapse
Role of NF-kB activation in cytolytic function

Key words: Natural killer cells, immunological synapse, cytotoxicity,
secretory lysosomes.

Description of Research
Natural killer (NK) cells are lymphocytes critical to host defense that play important roles in surveillance of tumor cells as well as in control of viral infections. They do not undergo genetic recombination to attain specificity and therefore are part of the innate immune system. NK cells mediate cytotoxicity by extruding secretory lysosomes in a directed manner after a favorable balance between the ligation of activating and inhibiting receptors has been achieved. The foundations of NK cell activities and regulation therefore lie at the interface between NK cells and cells with which they are interacting. Molecules accumulate in this region and result in a dynamic structure called the NK cell immunologic synapse (NKIS).

My laboratory is investigating the formation, function and regulation of the NKIS. We have focused upon the cytoskeleton as a critical juncture for these processes due to an interest in a human disease that impairs cytoskeletal function called the Wiskott-Aldrich syndrome. Using cells from patients with this disorder, as well as various cytoskeletal inhibitors, we have shown that the activating NKIS is actin-dependent. This is in contrast to the inhibitory NKIS, which is actin-independent. We have also identified sequential steps required for creation of the activating NKIS and have demonstrated that actin reorganization precedes and is required for microtubular function at the synapse. The microtubules are then needed to translocate lytic granules to the center of the NKIS called the central supramolecular activation cluster (cSMAC). Most recently our work has focused upon cytoskeletal events critical in forming the activating NKIS and we have been evaluating actin complex-associated proteins for their role in granule localization to the cSMAC. We are additionally studying how molecular rearrangement at the NKIS results in activation-induced transcriptional regulation and how this ultimately affects cytotoxic function. NK cells are a most useful model for these studies because they have an easily defined function and a more gradual activation process attributed to the interplay of activating and inhibitory receptors.

Rotation Projects for 2006-2007
1. Mechanism by which NF-kB essential modulator function and NF-kB activation enable cytolytic function
2. Biochemical and spatial evaluation of functional linkages between the actin cytoskeleton and the microtubular network in formation of the cytolytic immunological synapse.
3. Requirements for and behavior of secretory lysosome traffic to the cytolytic immunological synapse

Lab personnel:
Linda Monaco Shawver - Research Technician Level III
Raquel P. Deering - Research Technician Level II
Christine Destephan - Research Technician Level I
Pinaki P. Banerjee - Postdoctoral Fellow
Rahul Pandey - Postdoctoral Fellow
Eric Hanson - Postdoctoral Fellow

Appointments
Assistant Professor of Pediatrics at University of Pennsylvania School of Medicine (2003 – 2010)
Associate Professor of Pediatrics at University of Pennsylvania School of Medicine (2010 – 2012)
Education
M.D., Brown University (1997)
Ph.D., Pathobiology, Brown University (1996)
A.B., Biology, Brown University (1990)
Selected Publications
Rak Gregory D, Mace Emily M, Banerjee Pinaki P, Svitkina Tatyana, Orange Jordan S. Natural killer cell lytic granule secretion occurs through a pervasive actin network at the immune synapse.. PLoS biology. Vol 9(9) . 2011 Sep:e1001151.
Orange Jordan S, Roy-Ghanta Sumita, Mace Emily M, Maru Saumya, Rak Gregory D, Sanborn Keri B, Fasth Anders, Saltzman Rushani, Paisley Allison, Monaco-Shawver Linda, Banerjee Pinaki P, Pandey Rahul. IL-2 induces a WAVE2-dependent pathway for actin reorganization that enables WASp-independent human NK cell function.. The Journal of clinical investigation. Vol 121(4) . 2011 Apr:1535-48.
Pedroza Luis A, Kumar Vipul, Sanborn Keri B, Mace Emily M, Niinikoski Harri, Nadeau Kari, de Moraes Vasconcelos Dewton, Perez Elena, Jyonouchi Soma, Jyonouchi Harumi, Banerjee Pinaki P, Ruuskanen Olli, Condino-Neto Antonio, Orange Jordan S. Autoimmune regulator (AIRE) contributes to Dectin-1-induced TNF-a production and complexes with caspase recruitment domain-containing protein 9 (CARD9), spleen tyrosine kinase (Syk), and Dectin-1.. The Journal of allergy and clinical immunology. 2011 Sep.
Mentlik, A. N., Sanborn, K. B., Holzbaur, E. L., Orange, J. S.. Rapid lytic granule convergence to the MTOC in natural killer cells is dependent on dynein but not cytolytic commitment. Mol Biol Cell. Vol 21(13) . 2010:2241-56.
Boztug Kaan, Schmidt Manfred, Schwarzer Adrian, Banerjee Pinaki P, Díez Inés Avedillo, Dewey Ricardo A, Böhm Marie, Nowrouzi Ali, Ball Claudia R, Glimm Hanno, Naundorf Sonja, Kühlcke Klaus, Blasczyk Rainer, Kondratenko Irina, Maródi László, Orange Jordan S, von Kalle Christof, Klein Christoph. Stem-cell gene therapy for the Wiskott-Aldrich syndrome.. The New England journal of medicine. Vol 363(20) . 2010 Nov:1918-27.
Sanborn, K.B, Rak, G., Maru, S.Y., Demers, K., Difeo, A., Martignetti, J.A., Betts, M.R., Favier, R., Banerjee, P.P., Orange, J.S.. Myosin IIA associates with NK cell lytic granules to enable their interaction with F-actin and function at the immunological synapse. 2009. J. Immunol.. Vol 182. 2009 JUN:in press.
Orange, J.S.. Formation and function of the lytic NK cell immunological synapse.. Nature Rev. Immunol.. Vol 8. 2008:713-725.
Banerjee, PP, Pandey, R, Zheng, R, Suhoski, MM, Monaco-Shawver, L, Orange, JS. Cdc42-interacting protein-4 functionally links actin and microtubule networks at the cytolytic NK cell immunological synapse. Journal Of Experimental Medicine. Vol 204(10) . 2007 OCT:2305-2320.
Chang, JT, Palanivel, VR, Kinjyo, I, Schambach, F, Intlekofer, AM, Banerjee, A, Longworth, SA, Vinup, KE, Mrass, P, Oliaro, J, Killeen, N, Orange, JS, Russell, SM, Weninger, W, Reiner, SL. Asymmetric T lymphocyte division in the initiation of adaptive immune responses. Science. Vol 315(5819) . 2007 MAR:1687-1691.
Pandey, R, DeStephan, CM, Madge, LA, May, MJ, Orange, JS. NKp30 ligation induces rapid activation of the canonical NF-kappa B pathway in NK cells. Journal Of Immunology. Vol 179(11) . 2007 DEC:7385-7396.