Contribution of physiologists to the identification of the humoral component of immunity in the 19th century

The history of antimicrobial humoral immunity usually focuses on the works of the German school at the end of the 19th century, born in the tradition of chemistry and disinfection. Starting from an old quarrel of priority about serotherapy between Emil von Behring (1854–1917) and the French physiologists Charles Richet (1850–1935) and Jules Héricourt (1850–1938), we first confirm that the latter stated the principle of serotherapy in 1888 and put it into practice before the seminal Behring's article in 1890, observing several adverse effects of this new immunotherapy. We also find that researchers who can be considered heirs of the French school of Physiology founded by Claude Bernard (1813–1878) also investigated the field of humoral immunity in the 1870–1880s. Maurice Raynaud (1834–1881), Auguste Chauveau (1827–1917), and eventually Charles Richet applied the experimental method of Claude Bernard to the young field of microbiology, illustrating a movement called by Jacques Léonard “physiologization of the pasteurism.” However, the contribution of physiologists in this field started before Louis Pasteur, leading to the conclusion that physiologists and chemists synergistically contributed to the birth of bacteriology and immunology.     

2nd PEGS Annual Symposium on Antibodies for Cancer Therapy: April 30–May 1, 2012, Boston,USA

The 2nd Annual Antibodies for Cancer Therapy symposium, organized again by Cambridge Healthtech Institute as part of the Protein Engineering Summit, was held in Boston, USA from April 30^th to May 1^st, 2012. Since the approval of the first cancer antibody therapeutic, rituximab, fifteen years ago, eleven have been approved for cancer therapy, although one, gemtuzumab ozogamicin, was withdrawn from the market.  The first day of the symposium started with a historical review of early work for lymphomas and leukemias and the evolution from murine to human antibodies. The symposium discussed the current status and future perspectives of therapeutic antibodies in the biology of immunoglobulin, emerging research on biosimilars and biobetters, and engineering bispecific antibodies and antibody-drug conjugates. The tumor penetration session was focused on the understanding of antibody therapy using ex vivo tumor spheroids and the development of novel agents targeting epithelial junctions in solid tumors. The second day of the symposium discussed the development of new generation recombinant immunotoxins with low immunogenicity, construction of chimeric antigen receptors, and the proof-of-concept of ‘photoimmunotherapy’. The preclinical and clinical session presented antibodies targeting Notch signaling and chemokine receptors.  Finally, the symposium discussed emerging technologies and platforms for therapeutic antibody discovery.     

2nd PEGS Annual Symposium on Antibodies for Cancer Therapy

The 2nd Annual Antibodies for Cancer Therapy symposium, organized again by Cambridge Healthtech Institute as part of the Protein Engineering Summit, was held in Boston, USA from April 30^th to May 1^st, 2012. Since the approval of the first cancer antibody therapeutic, rituximab, fifteen years ago, eleven have been approved for cancer therapy, although one, gemtuzumab ozogamicin, was withdrawn from the market. The first day of the symposium started with a historical review of early work for lymphomas and leukemias and the evolution from murine to human antibodies. The symposium discussed the current status and future perspectives of therapeutic antibodies in the biology of immunoglobulin, emerging research on biosimilars and biobetters, and engineering bispecific antibodies and antibody-drug conjugates. The tumor penetration session was focused on the understanding of antibody therapy using ex vivo tumor spheroids and the development of novel agents targeting epithelial junctions in solid tumors. The second day of the symposium discussed the development of new generation recombinant immunotoxins with low immunogenicity, construction of chimeric antigen receptors, and the proof-of-concept of ‘photoimmunotherapy’. The preclinical and clinical session presented antibodies targeting Notch signaling and chemokine receptors. Finally, the symposium discussed emerging technologies and platforms for therapeutic antibody discovery.     

In the Cradle of Heredity; French Physicians and L'Hérédité Naturelle in the Early 19th Century

This paper argues that our modern concept of biological heredity was first clearly introduced in a theoretical and practical setting by the generation of French physicians that were active between 1810 and 1830. It describes how from a traditional focus on hereditary transmission of disease, influential French medical men like Esquirol, Fodéré, Piorry, Lévy, moved towards considering heredity a central concept for the conception of the human bodily frame, and its set of physical and moral dispositions. The notion of heredity as a natural force, with a wide ranging capabilities of transmitting differentially both fundamental and accidental characters was generalized by that generation of physicians with the help of contemporary naturalists and physiologists. By 1830 the term hérédité was widespread, and it shared the explanatory and semantic qualities of traditional medical concepts like constitution and temperament. An analysis is given of the main developments that led to the conception of biological (including human) bodies as consisting of a layered, hierarchical organization of characters, differentially affected by the law of conservation (Heredity) and change (Inneity, Variation). The mid-century work of the French physician Prosper Lucas, Traité Philosophique et Physiologique de L' Hérédité Naturelle, is shown to be the culmination of the efforts of several generations of French physicians towards having a feasible, complexly structured notion of how heredity works.     

Pioneers of exfoliative cytology in the 19th century: the predecessors of George Papanicolaou

The purpose of our study was to summarize the knowledge on exfoliative cytology during the 19th century and to track down Papanicolaou's predecessors. A thorough study of texts, medical books and reports, together with a review of the available literature in PubMed, was undertaken. The study of cytological preparations as a diagnostic procedure can be traced back to the work of the famous French microscopist Alfred François Donné. However, the systematic study and the criteria for the diagnosis of malignant cells should be attributed to Johannes Müller. The increasing interest in the cytological examination of various fluids of the human body can be confirmed by a plethora of studies published during this period. By the end of the 19th century, the invention of new techniques in pathology, such as the introduction of cell block techniques, tissue sections and new staining methods which provided the opportunity to study surgical specimens in three dimensions, led to a decrease in the interest in exfoliative cytology, which was re‐discovered by George Papanicolaou almost three decades later.     

Antibodies targeting G protein-coupled receptors: Recent advances and therapeutic challenges

Le STUDIUM conference was held November 24–25, 2016 in Tours, France as a satellite workshop of the 5^th meeting of the French GDR 3545 on “G Protein-Coupled Receptors (GPCRs) -From Physiology to Drugs,” which was held in Tours during November 22–24, 2016. The conference gathered speakers from academia and industry considered to be world leaders in the molecular pharmacology and signaling of GPCRs, with a particular interest in the development of therapeutic GPCR antibodies (Abs). The main topics were new advances and challenges in the development of antibodies targeting GPCRs and their potential applications to the study of the structure and function of GPCRs, as well as their implication in physiology and pathophysiology. The conference included 2 sessions, with the first dedicated to the recent advances in methodological strategies used for GPCR immunization using thermo-stabilized and purified GPCRs, and the development of various formats of Abs such as monoclonal IgG, single-chain variable fragments and nanobodies (Nbs) by in vitro and in silico approaches. The second session focused on GPCR Nbs as a “hot” field of research on GPCRs. This session started with discussion of the pioneering Nbs developed against GPCRs and their application to structural studies, then transitioned to talks on original ex vivo and in vivo studies on GPCR-selective Nbs showing promising therapeutic applications of Nbs in important physiologic systems, such as the central nervous and the immune systems, as well as in cancer. The conference ended with the consensus that Abs and especially Nbs are opening a new era of research on GPCR structure, pharmacology and pathophysiology.     

G6PD deficiency, diet, and adaptation to malaria

The symposium on “G6PD Deficiency, Diet, and Adaptation to Malaria” was held in Cortona, Italy on July 3,4,5 1995 under the auspices of the Congress of the International Union of Anthropological and Ethnological Sciences (IUAES). The Congress had actually taken place in Florence in April 1995, and the G6PD symposium was a satellite session to that meeting. Professor Brunetto Chiarelli, of the Istituto di Antropologia at the University of Florence, was the program chairman for the Congress. The general theme of the Congress was “Biodemography and Human Evolution,” and the G6PD symposium was consistent with this topic. In its broadest sense the symposium focused on biocultural factors which have influenced evolution at the G6PD locus and the pattern of population variation that has consequently emerged in this genetic system. A more specific sub-theme, reflected in the title of the symposium, was the interaction between dietary factors and the G6PD locus in providing antimalarial protection to human populations.     

Antibody modeling: implications for engineering and design

Our understanding of the rules relating sequence to structure in antibodies has led to the development of accurate knowledge-based procedures for antibody modeling. Information gained from the analysis of antibody structures has been successfully exploited to engineer antibody-like molecules endowed with prescribed properties, such as increased stability or different specificity, many of which have a broad spectrum of applications both in therapy and in research. Here we describe a knowledge-based procedure for the prediction of the antibody-variable domains, based on the canonical structures method for the antigen-binding site, and discuss its expected accuracy and limitations. The rational design of antibody-based molecules is illustrated using as an example one of the most widely employed modifications of antibody structures: the humanization of animal-derived antibodies to reduce their immunogenicity for serotherapy in humans.     

Therapeutic antibodies and infectious diseases,Tours, France,November 20−22, 2012

The Therapeutic Antibodies and Infectious Diseases international congress was held in Tours, France on November 20−22, 2012. The first session was devoted to the development of antibodies directed against bacterial toxins or viruses that could be used in a potential bioterrorist threat situation. The second session dealt with the effector functions of anti-microbial antibodies, while the third was oriented toward anti-viral antibodies, with a special emphasis on antibodies directed against the human immunodeficiency and hepatitis C viruses. After a lecture by a speaker from the US Food and Drug Administration on antibody cocktails, the second day ended with a special session dedicated to discussions regarding the involvement of French biotechnology industries in the field. On the last day, the congress concluded with talks about current antibody treatments for infectious diseases, with a particular focus on their adverse events. Participants enjoyed this very stimulating and convivial meeting, which gathered scientists from various countries who had different scientific research interests.     

2nd Charles Richet et Jules Héricourt Workshop: Therapeutic antibodies and anaphylaxis; May 31–June 1, 2011; Tours,France

The Charles Richet and Jules Héricourt workshops honor the memory of Jules Héricourt (1850–1938) and Charles Richet (1850–1935) who described the principle of serotherapy in 1888 and made the very first attempts to fight cancer with serotherapy in 1895. In 1902, Charles Richet and Paul Portier described “anaphylaxis,” a discovery awarded the Nobel Prize in 1913. The first workshop, “Towards the clinical use of monoclonal antibodies with higher cytolytic efficacy in cancer” was held in Tours, France on November 20–21, 2008. The second Charles Richet and Jules Héricourt workshop, held May 31–June 1, 2011 at the University of Tours, France, was also organized by the Cancéropôle Grand Ouest. The topic of the workshop was therapeutic antibodies and anaphylaxis, a subject rarely addressed in congresses focused on mAbs. To have discussions about mAb side effects with complete objectivity, the congress was organized independently of any sponsorship from pharmaceutical companies. This academic event was motivated by the high incidence of shocks to cetuximab and the need to compile and evaluate scattered information. This growing public health concern was thus analyzed from different scientific and medical angles. The first session was devoted to acute infusion reactions, with an emphasis on deconvolution of the terms “cytokine-release syndrome,” “cytokine storms,” “anaphylaxis” and their epidemiology. This session concluded with the Charles Richet lecture on cetuximab anaphylaxis and anti-αGal IgE by Thomas Platts-Mills, its discoverer. In the next session, the involvement of anti-glycan antibodies in both anaphylaxis and delayed hypersensitivity reactions to therapeutic antibodies was discussed. A gala dinner was held in the gardens of the beautiful château of Villandry, which was acquired and restored by Joachim Carvalho, a pupil of Charles Richet''s and great-grandfather of the present owner. The final session focused on strategies to prevent cetuximab anaphylaxis in clinical practice included a variety of topics, e.g., premedication, biobetters and biosimilars, skin testing and predictive assays. All speakers and attendees enjoyed this very stimulating and rewarding meeting, which gathered many people with divergent scientific backgrounds and medical specialties.Key words: cetuximab, anaphylaxis, α-Gal epitope, anti-αGal IgE, anti-cetuximab IgE, allergy, monoclonal antibodies, glycosylation     

From Anomaly to Unification: Tracy Sonneborn and the Species Problem in Protozoa, 1954--1957

This article examines the critique of the biological species concept advanced by protozoan geneticist Tracy Sonneborn at the 1955 AAAS symposium on “the species problem,” published subsequently in 1957. Although Sonneborn was a strong proponent of a population genetical conception of species, he became critical of the biological species concept for its failure to incorporate asexual and obligatory inbreeding organisms. It is argued that Sonneborn's intimate knowledge of the ciliate protozoan Paramecium aurelia species complex brought him into conflict with a growing pressure in the biological sciences to emphasize universal principles of life. Faced with the need to defend the value of P. aurelia as an investigative tool, Sonneborn argued that the sharp break in nature between sexual and asexual organisms posited by proponents of the biological species concept was not an existential feature of the living world, but rather the misleading consequence of an operational definition of species based only upon sexual organisms. Drawing upon his knowledge of the immense variability of P. aurelia, he proposed instead a continuum of breeding systems from obligatory outbreeding to asexual organisms, and a more broadly unifying definition of species that incorporated asexual as well as sexual organisms. Paradoxically, the push for unification that then characterized the evolutionary synthesis served to debar critical consideration of Sonneborn's more unificatory alternative, and his underlying contention that biological anomaly could serve as an important source of conceptual unification. This revised version was published online in July 2006 with corrections to the Cover Date.     

Using phylogenies in conservation: new perspectives

The 2011 meeting of the European Ecological Federation took place in ávila, Spain, from 26th September to 29th September. The French Ecological Society (SFE) and the Foundation for Research on Biodiversity (FRB) sponsored a session entitled 'Evolutionary history, ecosystem function and conservation biology: new perspectives'. We report on the main insights obtained from this symposium.     

Production,characterization, and immunocytochemical applications of monoclonal antibodies to human sperm protamines

Three monoclonal antibodies against human protamines were obtained by immunization with total human basic nuclear proteins or purified protamine HP3. The specificity of antibodies was assessed by enzyme-linked immunosorbent assay (ELISA) and Western blot. They recognized three distinct epitopes: One was specific for the protamine P1 family, another was specific for the protamine P2 family and the third was common to both families. All were specific for the human species. Antibodies were used to detect protamines in germ cells by indirect immunofluorescence and by immunoelectron microscopy. Protamines appeared in spermtid nuclei at steps 4–5 of spermiogenesis, i.e., during the chromatin condensation process, and were not accumulated in the cytoplasm before entering the nucleus. © 1993 Wiley-Liss, Inc.     

MAbDelivery: Administration routes for antibody therapy Third LabEx MAbImprove industrial workshop,July 2, 2015 Tours,France

The annual “LabEx MAbImprove Industrial Workshops” are primarily intended to provide a comprehensive view about topics of interest for the pharmaceutical industry to scientists involved in research on therapeutic antibodies. The third workshop in this series, held July 2, 2015 in Tours, was dedicated to the optimization of delivery, namely all processes leading monoclonal antibodies to reach their target site. The commonly used intravenous (IV) route, although advantageous in terms of pharmacokinetics and pharmacodynamics, presents some disadvantages in terms of patients' convenience, therapeutic target access or treatment cost. Such problems led pharmaceutical companies to consider more straightforward and patient-friendly administration routes, bringing the need for specific formulations adapted to the specific inherent physicochemical challenges. In this context, the workshop provided an overview of these advances and opened discussion on new administration routes and formulation development. In the first session, the opportunities and challenges of 3 main routes of administration (IV, subcutaneous (SC), and pulmonary) were discussed, integrating protein stability issues. The next session was dedicated to medical devices intended for SC and pulmonary administration. The last session focused on specific formulations for monoclonal antibodies, particularly to successfully protect antibodies upon aerosolization, to develop highly concentrated formulations for SC administration, and to use formulation as a mean to overcome the barriers to oral protein delivery. As in the previous editions, this workshop gathered people from the academic and industrial spheres and allowed rich debates and discussions.     

Stature variation in the British American Colonies: French and Indian War records, 1755-1763

Personnel records kept by military units of American colonials during the French and Indian War (1755-1763) are analyzed for relationships between environmental factors and stature. A robust American economy and direct access to high-quality food were apparently critical to tallness of this white American male sample. American-born men were taller at all ages than those who had migrated from Europe. January temperatures, rural versus urban birth, and ethnicity also showed stature relationships within the American-born group; thermal effects were by far the strongest of the non-nutritional factors.     

Multifunctionalization of cetuximab with bioorthogonal chemistries and parallel EGFR profiling of cell-lines using imaging,FACS and immunoprecipitation approaches

The ability to derivatize antibodies is currently limited by the chemical structure of antibodies as polypeptides. Modern methods of bioorthogonal and biocompatible chemical modifications could make antibody functionalization more predictable and easier, without compromising the functions of the antibody. To explore this concept, we modified the well-known anti-epidermal growth factor receptor (EGFR) drug, cetuximab (Erbitux®), with 5-azido-2-nitro-benzoyl (ANB) modifications by optimization of an acylation protocol. We then show that the resulting ANB–cetuximab can be reliably modified with dyes (TAMRA and carboxyrhodamine) or a novel synthesized cyclooctyne modified biotin. The resulting dye- and biotin-modified cetuximabs were then tested across several assay platforms with several cell lines including U87, LN229, F98_EGFR, F98_WT and HEK293 cells. The assay platforms included fluorescence microscopy, FACS and biotin–avidin based immunoprecipitation methods. The modified antibody performs consistently in all of these assay platforms, reliably determining relative abundances of EGFR expression on EGFR expressing cells (LN229 and F98_EGFR) and failing to cross react with weak to negative EGFR expressing cells (U87, F98_WT and HEK293). The ease of achieving diverse and assay relevant functionalizations as well as the consequent rapid construction of highly correlated antigen expression data sets highlights the power of bioorthogonal and biocompatible methods to conjugate macromolecules. These data provide a proof of concept for a multifunctionalization strategy that leverages the biochemical versatility and antigen specificity of antibodies.     

Using drug-discrimination techniques to study the abuse-related effects of psychoactive drugs in rats

Drug-discrimination (DD) techniques can be used to study abuse-related effects by establishing the interoceptive effects of a training drug (e.g., cocaine) as a cue for performing a specific operant response (e.g., lever pressing reinforced by food). During training with this protocol, pressing one lever is reinforced when the training drug is injected before the start of the session, and responding on a second lever is reinforced when vehicle is injected before the session. Lever choice during test sessions can then be used as an indication of whether a novel drug has effects similar to the training drug, or whether a potential therapeutic alters the effects of the training drug. Although training can be lengthy (up to several months), the pharmacological specificity of DD procedures make them a perfect complement to other techniques used to study drug-abuse phenomena, such as intravenous self-administration and conditioned place-preference procedures.     

Recognition of riboflavin and the capsular polysaccharide of Haemophilus influenzae type b by antibodies generated to the haptenic epitope D-ribitol

D-Ribitol, a five–carbon sugar alcohol, is an important metabolite in the pentose phosphate pathway; it is an integral part of riboflavin (vitamin B2) and cell wall polysaccharides in most Gram-positive and a few Gram-negative bacteria. Antibodies specific to D-ribitol were generated in New Zealand white rabbits by using reductively aminated D-ribose-BSA conjugate as the immunogen. MALDI-TOF and amino group analyses of ribitol-BSA conjugate following 120 h reaction showed ~27–30 mol of ribitol conjugated per mole BSA. The presence of sugar alcohol in the conjugates was also confirmed by an increase in molecular mass and a positive periodic acid–Schiff staining in SDS-PAGE. Caprylic acid precipitation of rabbit serum followed by hapten affinity chromatography on ribitol–KLH–Sepharose CL-6B resulted in pure ribitol–specific antibodies (~45–50 μg/mL). The affinity constant of ribitol antibodies was found to be 2.9?×?10⁷ M^?1 by non-competitive ELISA. Ribitol antibodies showed 100 % specificity towards ribitol, ~800 % cross–reactivity towards riboflavin, 10–15 % cross–reactivity with sorbitol, xylitol and mannitol, and 5–7 % cross–reactivity with L-arabinitol and meso-erythritol. The specificity of antibody to ribitol was further confirmed by its low cross-reactivity (0.4 %) with lumichrome. Antibodies to D-ribitol recognized the purified capsular polysaccharide of Haemophilus influenzae type b, which could be specifically inhibited by ribitol. In conclusion, antibodies specific to D-ribitol have been generated and characterized, which have potential applications in the detection of free riboflavin and ribitol in biological samples, as well as identification of cell-surface macromolecules containing ribitol.     

Serotherapy of ovarian cancer

The development of monoclonal antibodies has permitted the identification of several ovarian-tumor-associated antigens which might serve as targets for serotherapy in vivo. With the exception of antibodies directed against growth factor receptors, unmodified monoclonal reagents must activate complement (C') components or bind effector cells to destroy tumor targets. Antibody-dependent cell-mediated cytotoxicity (ADCC) may be particularly important for eliminating tumor cells in vivo. A shortage of functionally active effector cells can limit the efficacy of serotherapy with heteroantisera or monoclonal reagents. The use of immunostimulants such as Corynebacterium parvum has increased the number and activity of effector cells for ADCC within the peritoneal compartment of mice and of patients with ovarian cancer. Intraperitoneal serotherapy can achieve direct contact between antibody and microscopic deposits of ovarian tumor cells which persist following cytoreductive operations and cytotoxic chemotherapy. Conjugation of monoclonal antibodies with radionuclides, drugs or toxins might increase the potency of serotherapy and circumvent the effector shortage. Clinical studies to date have evaluated radionuclide conjugates for imaging and for therapy. Patients with a small volume of disease have responded to treatment. Preclinical models suggest that drug and toxin conjugates might also prove active. Recent studies have demonstrated a synergistic interaction between different immunotoxins. Ovarian carcinoma is likely to be a valuable clinical model for evaluating immunoconjugates which react with epithelial tumor cells.