Manufacturing complex biologics for 

regenerative immunology

J H Elisseeff

Morton Goldberg Professor

Johns Hopkins University

December 1, 2022

@JHElisseeff
TISSUE ENGINEERING AS A STRATEGY FOR REPAIR

Biomaterial scaffolds + biological signals + cells

Clinical translation slow in tissue engineering

Global organ donation:

136K, >7% yearly growth

Osteoarthritis:

Single most common cause 

of disability >60, ~10-15% of 

population
Lessons in translation: Regenerative immunotherapies for tissue repair

gd

B

ILC

CD4+

Gran

Mac

Mono

IFNg

IL4, IL17

M1, M2

Understand the biomaterial immune response and engineer for repair

CLINICAL TRANSLATION INFORMS NEW RESEARCH DIRECTION

Cartilage Repair

Soft Tissue Filler

Redirecting wound healing

Tissue specific immune response
REGENERATIVE IMMUNOLOGY

Marrying regenerative medicine, immunology and tissue engineering

Define immune impact 

on tissue development

The immune environment 

determines downstream 

tissue repair

Design and apply

Regenerative

Immunotherapies

The immune system is therapeutically accessible → the right target for regenerative medicine

Engineer immunotherapies 

to create a proregenerative environment

Map the immune response 

to injury (and biomaterials)

The immune system is 

the first responder to 

injury 

Innate Adaptive

T cells

Neu Mac B cells

ILC

Th2 T cells

Senescence

Stem cells
www.aestheticplasticsurgerypc.com Badylak SF et al. Tiss Eng Part A. 2015.

Breast

Esophagus

Dura

Biodesign® Dural Graft

Surgisis® DurepairTM Regeneration Matrix

AlloDermTM

PermacolTM

BIOLOGICAL SCAFFOLDS IN THE CLINIC

Minimally processed extracellular matrix (ECM)

ECM materials contain DAMPs and intracellular proteins
MUSCLE AND BIOLOGICAL SCAFFOLDS: PRO-REGENERATIVE IMMUNE PROFILE

T cells in the Wound/biomaterial

IL4 Expression

1 0

1 0 0

1 0 0 0

re a te d (2

-D D C t

)

****

aline (2-ddCt

)

Absolute Cell Number

A b s o lu te C e ll N u m b e r

A

C

IL-4 RQ to Saline (2-ddCt

)

Distal Local

Lymph nodes

B

D

Lymph node changes

500 μm

200 μm

S

alin

e

C-E

C

M

500 μm

200 μm

Saline

C-EC

M

Implant

Control

Biomaterials ENHANCE the TH2 response and create a pro-regenerative environment in muscle

Sadtler et al., Science, 2016

T cells direct 

macrophage behavior

SYSTEMIC IMPACT
ECM: composition and biologic activity consistency

• Physical properties

• Color, water, DSC, other

• Specific components as metric - lipids, collagen?

• Mechanics/Injectability

• Biological activity

• Promote appropriate cell infiltration

• Stimulate new tissue development

ECMs are complex biologics and should be consistently regulated 
IL-1b

TYPE 2 IMMUNE RESPONSE: target for regenerative medicine
TYPE 2 IMMUNE STIMULATION

Inspiration from nature → Schistosomiasis soluble egg antigen

Maestas et al, Biorvx, 2022
SEA: FORMULATING FOR REPAIR AND REGENERATION

Maestas et al, Biorvx, 2022
Il4 Il13 Il23a Ifng

-4

0

4

8

1wk Muscle RQ to Saline Log2(FC)

Saline SEA

rSEA

✱✱

✱✱✱✱

✱✱✱✱

✱✱✱✱

✱✱✱✱

✱

✱✱

✱✱

1d 3d 1wk 2wk 3wk 6wk

-2

0

2

4

6

iLN Il4 Gene Exp. RQ to SalineLog2

(FC)

 

** **

***

rSEA PROMOTES EXTENDED TYPE 2 IMMUNE RESPONSE

Maestas et al, Biorvx, 2022
rSEA PROMOTES EXTENDED TYPE 2 IMMUNE RESPONSE

Maestas et al, Biorvx, 2022

1wk 3wk

-3

0

3

6

9

Il4 Gene Exp. Muscle

RQ to SalineLog2

(FC)

Saline

rSEA

Vitrigel Vitrigel+rSEA

✱ ✱

✱✱✱

✱✱✱✱

✱✱✱

Saline

Vitrigel rSEA-Vitrigel

rSEA

rSEA-Vitrigel

Digested ECM rSEA

ECM-Hydrogel

Vitrification

Vitrigel
rSEA: A COMPLEX MIXTURE

Proteins

Lipids

SEA

rSEA
Validating biological activity: developing relevant screens
From lessons in translation to

Lost in translation...

Microbiome Infection

Aging

Cancer 

(comorbidities)
Dystrophin / DAPI / Laminin

REGENERATIVE BIOMATERIAL RESPONSE IN AGING ANIMALS?

Increased adipogenesis and 

fibrosis in 20 month old mice

Reduced muscle repair

Han, Biorvx, 2021
Aging

Need for combination therapies

Han et al, Biorvx, 2022
ADAPTIVE IMMUNE CHANGES WITH AGING

Han, Biorvx, 2021

gd T cells CD4 T cells
Il17f Il17a

Ifng

INJURY UNCOVERS AGING DIFFERENCES IN 

SECONDARY IMMUNE STRUCTURES 
IMMUNE CHANGES IN AGING MUSCLE INJURY

Han, Biorvx, 2021

Reparative 

immune cells 

TH17 

gd

TH2

Eosinophils

CD8+

Regenerative immune cells decrease and inflammatory cells increase with age Senescent cells: a stromal bridge to the immune system
IL-1b

IL17 negative regulation of IL4
COMBINATION THERAPY RESTORES TISSUE REPAIR

ECM + Isotype

200μm

Isotype

1mm

ECM + αIL17A

VML 7d

Old 4Get

• IL-17A or F antibody (αIL17A or F)

• 10μg antibody in 20μL volume

• Intra-muscular (IM) injection x 3

9d 11d 3 wk

0

20

40

60

80

IL4+ Eosinophil # in Mu

(x10 count)

**

 

** **

αIL17A

αIL17F

Isotype

No injury

0

20

40

60

IL4+ CD4 T # in Mu

(x10 count)

**

*

αIL17A

αIL17F

Isotype

No injury
Manufacturing complex biologics

• Needed for targeting the multiple contributors to repair

• Multiple mechanisms of action

• Consistent regulation needed

• Physical, compositional and biological release assays

• Combination products for aging and personalized medicine
Acknowledgements

Bloomberg~Kimmel Instititue for Cancer 

Immunotherapy

Drew Pardoll

Jonathan Powell

Franck Housseau

Sudipto Ganguly

Ada Tam

Janis Taube, Bob Anders

Current Lab members

Chris Cherry Kavita Krishnan

David Maestas Brenda Yang

Anna Ruta Alex Chin

Joscelyn Meijas Helen Nguyen

Josh Hooks Locke Davenport Huyer

Alexis Pena Katlin Stivers

Jin Han Alexis Pena

John Michel

@JHElisseeff

Recent Lab Alumni

Liam Chung

Heather Jacobs

Kaitlyn Sadtler

Okhee Jeon

Chaekyu Kim

Matthew Wolf, PhD

Sven Sommerfeld, PhD

Jim Andorko

Hong Zhang

Jordan Garcia, MD

Erika Moore

Senescence 

Judy Campisi, Buck Institute

Darren Baker, Mayo

Jan van Deursen

Doahong Zhou, UF

Computational

Elana Fertig

Patrick Cahan

Chris Cherry

Clinical

Gedge Rosson, Plastic Surgery

Clifton (Bing) Bingham, Rheum

Damon Cooney, Plastic Surgery

Gerald Brandacher, Plastic Surgery

TTEC Administration 

Bahar Zarrabi, MBA

Dallas Auer

Pam Goldberg

Eileen Snyder
Aged T cells differentiate more to Th17 and secrete different proteins

AGED T CELLS ARE DIFFERENT

Han, Biorvx, 2021
AGED SnCs ARE DIFFERENT IN MUSCLE WOUND

Young and old senescent cells are a little different!

Jin Han

Senescent cells