{"doi":"10.1155/2022/6993187","title":"Synthesis, Characterization, and In Vivo Cytokinome Profile of IL-12-Loaded PLGA Nanospheres","abstract":"We report the successful encapsulation and elution of recombinant murine IL-12 (rmIL-12) from poly(lactide-co-glycolic) acid (PLGA) nanospheres (IL-12-NS) synthesized using the double emulsion/solvent evaporation (DESE) technique with microsphere depletion through ultracentrifugation. Images obtained with scanning electron microscopy (SEM) showcased a characteristic spherical shape with a mean particle diameter of <a:math xmlns:a=\"http://www.w3.org/1998/Math/MathML\" id=\"M1\"><a:mn>138.1</a:mn><a:mo>±</a:mo><a:mn>10.8</a:mn><a:mtext> </a:mtext><a:mtext>nm</a:mtext></a:math> and zeta potential of <c:math xmlns:c=\"http://www.w3.org/1998/Math/MathML\" id=\"M2\"><c:mo>−</c:mo><c:mn>15.1</c:mn><c:mo>±</c:mo><c:mn>1.249</c:mn><c:mtext> </c:mtext><c:mtext>mV</c:mtext></c:math> . These values suggest minimal flocculation when in solution, which was reflected in an in vivo biodistribution study that reported no observed morbidity/mortality. Encapsulation efficiency (EE) was determined to be <e:math xmlns:e=\"http://www.w3.org/1998/Math/MathML\" id=\"M3\"><e:mn>0.101</e:mn><e:mo>±</e:mo><e:mn>0.009</e:mn><e:mi>%</e:mi></e:math> with average particle concentration obtained per batch of <g:math xmlns:g=\"http://www.w3.org/1998/Math/MathML\" id=\"M4\"><g:mn>1.66</g:mn><g:mo>×</g:mo><g:msup><g:mrow><g:mn>10</g:mn></g:mrow><g:mrow><g:mn>9</g:mn></g:mrow></g:msup><g:mo>±</g:mo><g:mn>4.45</g:mn><g:mo>×</g:mo><g:msup><g:mrow><g:mn>10</g:mn></g:mrow><g:mrow><g:mn>8</g:mn></g:mrow></g:msup></g:math> particles/mL. Disparate zeta (ζ) potentials obtained from both protein-loaded and protein-unloaded batches suggested surface adsorption of protein, and confocal microscopy of BSA-FITC-loaded nanospheres confirmed the presence of protein within the polymeric shell. Furthermore, elution of rmIL-12 from IL-12-NS at a concentration of 500 million particles/mL was characterized using enzyme-linked immunosorbent assay (ELISA). When IL-12-NS was administered in vivo to female BALB/c mice through retroorbital injection, IL-12-NS produced a favorable systemic cytokine profile for tumoricidal activity within the peripheral blood. Whereas IFN-γ nadir occurred at 72 hours, levels recovered quickly and displayed positive correlations postburst out to 25 days postinjection. IL-12-NS administration induced proinflammatory changes while prompting minimal counterregulatory increases in anti-inflammatory IL-10 and IL-4 cytokine levels. Further, while IL-6 levels increased to 30 folds of the baseline during the burst phase, they normalized by 72 hours and trended negatively throughout the sill phase. Similar trends were observed with IL-1β and CXCL-1, suggesting a decreased likelihood of progression to a systemic inflammatory response syndrome-like state. As IL-12-NS delivers logarithmically lower amounts of IL-12 than previously administered during human clinical trials, our data reflect the importance of IL-12-NS which safely create a systemic immunostimulatory environment.","journal":"Journal of Immunology Research","year":2022,"id":280361,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":10,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9646,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2022-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":588034,"name":"Justin Markel","orcid":"0000-0003-2244-5185","position":1,"is_corresponding":false},{"id":955301,"name":"Jabeen Noore","orcid":null,"position":2,"is_corresponding":false},{"id":663849,"name":"Hillary G. Pratt","orcid":"0000-0002-3058-7434","position":3,"is_corresponding":false},{"id":440453,"name":"Brock A. Lindsey","orcid":"0000-0002-1073-5372","position":4,"is_corresponding":false},{"id":954797,"name":"Ryan A. Lacinski","orcid":"0000-0002-3026-2592","position":0,"is_corresponding":true}],"reference_count":74,"raw_metadata":null,"created_at":"2026-07-19T00:29:03.247347Z","pmid":"35465347","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}